Methods of inhibiting the ghrelin/growth hormone secretatogue receptor pathway and uses thereof

ABSTRACT

The invention provides methods for treatment, prevention or management of obesity, obesity related disorders, diabetes mellitus, and metabolic syndrome in a subject by administering a ghrelin O-acyltransferase (GOAT) inhibitor and/or a ghrelin receptor antagonist to the subject. The invention also provides ghrelin receptor antagonists of formula (VII): A 11 -A 12 -A 13 -Gly-Ser-A 14 -Phe-Leu-A 15 -A 16 -A 17 -A 18 , wherein each of A 11 , A 12 , and A 13  is independently absent, an amino acid, or an amino protecting group; each of A 15 , A 16 , A 17 , and A 18  is independently absent or an amino acid; and A 14  is a serine conjugated with a —C(O)C 1 -C 20 alky or a diaminopropionic acid conjugated with a —C(O)C 1 -C 20 alkyl group, provided that at least one of A 11 , A 12 , or A 13  is present.

CROSS REFERENCE TO RELATED APPLICATIONS

This application is a Continuation of U.S. patent application Ser. No.13/320,408, filed on Feb. 3, 2012, which is a 35 U.S.C. §371 NationalPhase Entry Application of International Application No.PCT/US2010/034570, filed May 12, 2010, which designates the UnitedStates, and which claims benefit of U.S. provisional application No.61/177,400, filed May 12, 2009, the content of which is hereinincorporated by reference in its entirety.

FIELD OF INVENTION

This invention relates to novel methods for treatment, management andprevention of a range of medical conditions that are associated with theactivation of the ghrelin/growth hormone receptor pathway. The medicalconditions include but are not limited to, obesity andobesity-associated disorders, diabetes, metabolic and/or endocrinedisorders, gastrointestinal disorders, cardiovascular disorders, centralnervous system disorders, genetic disorders, and hyperproliferativedisorders. Methods of modulating of the ghrelin/growth hormone receptorpathway include simultaneously inhibiting the synthesis of activeghrelin and inhibiting ghrelin receptor pathway activation Inhibition ofthe synthesis of active ghrelin is achieved by a ghrelinO-acyltransferase (GOAT) inhibitor. Inhibition of ghrelin receptoractivation is achieved by ghrelin receptor antagonists or inverseagonist.

BACKGROUND OF THE INVENTION

In America, there is a dramatic raise in the number of people who areoverweight. About 66% of U.S. adults age 20 or older are overweight.About 31% of American adults are obese today compared to 24% in 1994.Childhood obesity is also fast increasing. The prevalence of overweighthas steadily increased over the years among both genders, all ages,racial and ethnic groups, educational levels, and smoking levels.Accompanying overweight and obesity are a host of life threateningdiseases-diabetes, heart disease, stroke, high blood pressure andcertain cancers—which greatly decrease the quality of life and shortenthe life expectancy of an overweight/obese person. Every year, tens ofthousands of severely obese individuals undergo some sort of bariatricsurgery to control their eating and weight problem. Weight loss andexercise programs are multi-million dollar industry in America. Millionsare also spent on treating the overweight and obesity-related diseases.Therefore, additional new approaches to addressing the overweight andobesity issue are useful and urgently needed in our modern society.

The growth hormone (GH) secretatogue receptor (GHS-R) (also known as theghrelin receptor) pathway is a powerful stimulator of pulsatile GHsecretion and the pathway exhibits intricate interactions with severalprimary hypothalamic GH regulators. The elevation of growth hormone (GH)levels in animals, e. g., mammals including humans, upon administrationof GH-releasing compounds can lead to enhanced body weight and metabolicdiseases/disorders such as obesity and diabetes mellitus. Examples ofGH-releasing compounds include ghrelin, an endogenous ligand for GHS-R,growth hormone releasing peptides (GHRPs) and growth hormonesecretatogues (GHSs), all of which are potent orexigenic peptides.Orexigenic compounds stimulate appetite. In the absence of the ghrelinreceptor, transgenic female and male mice fed a high fat diet eat lessfood, less of the consumed calories are stored, fat is more of theenergy substrate, and body weight and body fat are less in these micethan control mice. When the ghrelin receptor was absent and mice werefed a normal diet, body weight and body fat were decreased in female butnot in male mice. In the absence of the ghrelin peptide, transgenic malemice (female mice not studied) had less rapid body weight gain on a highfat diet. This was associated with increased energy expenditure andincreased locomotive activity as well as decreased adiposity. Thesestudies indicate the ghrelin pathway is involved in body weight controlespecially when consuming a high caloric type of obese inducing diet. Inthe absence of the ghrelin receptor (GHS-R 1a), ghrelin no longerincreased food intake. Thus, the singularity of this receptor formediating ghrelin induced food intake is indicated. Also, hyperphagia isan established risk factor in diabetes mellitus in humans and evidenceindicates that sub-threshold doses of ghrelin increases food intake instreptozotocin treated rats. Experimental studies in rats revealedinterrelationships of ghrelin, somatostatin and GHRH on function of theGH axis.

Plasma levels of ghrelin rise precipitously in the blood before meals,when the stomach is empty, and fall after or during food consumption.Since intracadiac venous (i.v.) or intracerebroventricular (i.c.v)administration of ghrelin increases food intake, it appears that thephysiological role of ghrelin is a link or messenger between the stomachand the hypothalamus and the pituitary. One hypothesis is that when anorganism is getting ready for a meal, the CNS sends signals to the GItract telling that a meal is about to be consumed in order to obtaininformation back about the status of the digestive process, state ofdistension etc. from the various chemical and mechanical sensors in thegut. Here, ghrelin could be an important hormonal messenger, which issent back to the central nervous system (CNS) as a signal telling thatthere is no food in the stomach and that the gastrointestinal (GI) tractis ready for a new meal. In such a paradigm it is clear that a blockerof the ghrelin receptor would be a very efficient anti-obesity agent, asit would block the meal initiating, appetite signal from the GI tract.

The ghrelin receptor, GHS-R 1a, belongs to a relatively small family of7 transmembrane G-protein coupled receptors. A number of findingsdemonstrate how the ghrelin receptor uniquely play a role in mediatingthe action on GH release and food intake. This includes ghrelin receptorgenetics, mutations, structure, intracellular signaling, highconstitutive activity, enhancement of the number of hypothalamic ghrelinreceptors during starvation, etc. A spectrum of growth and metabolicchanges occur in mice as a result of knockout of the ghrelin molecule aswell as the ghrelin receptor. Adiposity in mice followed over expressionof the ghrelin receptor in hypothalamic growth hormone releasing hormone(GHRH) arcuate neurons. Over time, select biological effects ofghrelin/GHSs, especially non-endocrine effects, have been revealed whichpresumably occur via subtypes receptors of ghrelin or perhaps ghrelinreceptors with select mutations. Evidence indicates binding andactivation of the multifunctional CD36 receptor by GHSs. Anothernoteworthy finding of the ghrelin receptor was that underpathophysiological conditions the density of this receptor was reportedto be five times greater in atherosclerotic coronary arteries.

Peptide antagonists that inhibited the binding activity of GHSs inhypothalamic tissue in vitro have been reported. This included thesubstance P (“Sub P”) analog, [DArg¹ DPhe⁵ DTrp^(7,9) Leu¹¹]-SubstanceP, that subsequently was demonstrated by Holst et al (J. Biol. Chem.282, 15799 (2007)) to have both inverse agonist and ghrelin-R antagonistactivity.

The ghrelin receptor, GHS-R 1a is a constitutively active receptor, i.e. there is spontaneous, ligand-independent signaling from thisreceptor. This constitutive activity can be inhibited by [DArg¹, DPhe⁵,DTrp^(7,9) Leu¹¹]-substance P analog which has been previouslycharacterized both in vitro and in vivo as a weak competitive receptorantagonist to acute and chronic actions of GHRP-2 and ghrelin. This SubP analog has 2 types of ghrelin receptor inhibiting activities. At a lowdose (5 nM, IC₅₀), this Sub P analog is a potent inverse receptoragonist since it decreases elevated intracellular IP3 levels in theabsence of ghrelin but also it is a weak ghrelin GHRP-6 competitivereceptor antagonist since high dosages (630 nM, IC₅₀) inhibited receptorbinding of both peptides. Continuous i.c.v 7 day infusion of a very lowdose of the Sub P ghrelin receptor inverse agonist inhibited body weightgain of male rats. This was a dose that would be too low to function asa competitive ghrelin receptor antagonist and thus it was considered tobe due to the inverse agonist activity of the Sub P analog. In vitroevidence supports GHS-R antagonists with only inverse agonist or onlyghrelin/GHS-R activity or a combination of the two.

It is proposed that the high constitutive activity of the ghrelinreceptor plays a key functional role at CNS sites at which the receptoris expressed within the blood brain barrier and thus does not haveimmediate access to circulating ghrelin. This is in contrast to theghrelin receptor located in the arcuate nucleus and dorsal vagal complexrole. Thus it is possible that select GHSs, because of their differentchemistry, may have ready access to brain sites inaccessible to ghrelin.If this occurs, GHSs' actions at these sites may alter the CNS ghrelinconstitutive activity via receptor number and/or activity.

Accordingly, methods that inhibit and/or disrupt the activity of theGHS-R signaling pathway are useful in regulating GH secretion, appetite,and body weight. Furthermore, since metabolic diseases and disorderssuch as obesity, diabetes mellitus, and inhibition of growth hormonesecreted from tumors such as pituitary, prostate, osteoblast, pancreaticand hepatoma are directly and indirectly associated with activities ofthe GH axis, new strategies that inhibit/disrupt this pathway'sactivation, particularly at the level of the CNS, are useful in thetreatment of these metabolic diseases and disorders and cancers.

SUMMARY OF THE INVENTION

The present invention provides novel strategies for inhibiting and/ordisrupting the activities of ghrelin and/or the ghrelin/GHS-R 1asignaling pathway in vivo for the treatment of metabolic diseases anddisorders such as obesity, overeating, diabetes mellitus, unregulatedcell proliferation and for the inhibition of growth hormone secretedfrom tumors such as pituitary, prostate, medullary thyroid carcinomas,osteoblast, pancreatic and hepatoma. The strategy is to simultaneouslyinhibiting the synthesis of active ghrelin and inhibiting ghrelinreceptor activation, and thus the ghrelin/GHS-R 1a signaling pathway. Insome embodiments, the strategy comprises inhibiting the synthesis ofactive ghrelin only Inhibition of the synthesis of active ghrelin isachieved by a ghrelin O-acyltransferase (GOAT) inhibitor. Inhibition ofghrelin receptor activation is achieved by a ghrelin receptor antagonistor inverse agonist. Examples of GOAT inhibitors and ghrelin receptorantagonists and inverse agonist are described herein.

Accordingly, provided herein is a method for inhibiting and/ordisrupting the GHS-R signaling pathway in a subject in need thereof, themethod comprising administering an effective amount of a ghrelinO-acyltransferase (GOAT) inhibitor and an effective amount of a ghrelinreceptor antagonist and/or inverse agonist. In one embodiment, providedherein is a method for inhibiting and/or disrupting the GHS-R signalingpathway in a subject in need thereof, the method comprisingadministering an effective amount of a GOAT inhibitor. In anotherembodiment, provided herein is a method for inhibiting and/or disruptingthe GHS-R signaling pathway in a subject in need thereof, the methodcomprising administering an effective amount of ghrelin receptorantagonist and/or inverse agonist. Such individuals can be one withovereating disorder, obesity, obesity related disease or disorder,diabetes mellitus, metabolic syndrome and cancer. It is alsocontemplated that a plurality of GOAT inhibitors and/or ghrelin receptorantagonists and/or inverse agonists are administered.

Accordingly, provided herein is a method for treatment, prevention ormanagement of obesity in a subject in need thereof, the methodcomprising the step of administering an effective amount of a ghrelinO-acyltransferase (GOAT) inhibitor and an effective amount of a ghrelinreceptor antagonist and/or inverse agonist.

In another embodiment, the invention provides a method for treatment,prevention or management of obesity related disease or disorder in asubject in need thereof, the method comprising the step of administeringan effective amount of a ghrelin O-acyltransferase (GOAT) inhibitor andan effective amount of a ghrelin receptor antagonist and/or inverseagonist. Obesity related disease or disorder can include but are notlimited to diabetes, hypertension, and metabolic syndrome.

In one embodiment, the method for treatment, prevention or management ofobesity and obesity related disease or disorder in a subject in needthereof further comprises an anti-obesity treatment. In one embodiment,the anti-obesity treatment is a combination of dietary restrictiontherapy with a 5HT (serotonin) transporter inhibitor, a NE(norepinephrine) transporter inhibitor, a CB-1 (cannabinoid-1)antagonist/inverse agonist, a H3 (histamine H3) antagonist/inverseagonist, a MCH1R (melanin concentrating hormone 1R) antagonist, a MCH2Ragonist/antagonist, a NPY1 antagonist, a leptin, a leptin derivative, aleptin analog, PYY(3-36), PYY(1-36), an opioid antagonist, an orexinantagonist, a BRS3 (bombesin receptor subtype 3) agonist, a CCK-A(cholecystokinin-A) agonist, a CNTF (Ciliary neurotrophic factor), aCNTF derivative, or a lipase drug inhibitor administered simultaneously,concurrently or sequentially. In another embodiment, the anti-obesitytreatment is bariatric surgery. In another embodiment, the anti-obesitytreatment is a physical exercise program.

In one embodiment, the invention provides a method for treatment,prevention or management of diabetes mellitus in a subject in needthereof, the method comprising the step of administering an effectiveamount of a GOAT inhibitor and/or an effective amount of a ghrelinreceptor antagonist and/or inverse agonist. The diabetes mellitus can betype I or II.

In another embodiment, the invention provides a method for themodulation of ghrelin receptor in a subject in need thereof, the methodcomprising the step of administering an effective amount of a GOATinhibitor and an effective amount of a ghrelin receptor antagonistand/or inverse agonist. In one embodiment, the invention provides amethod for the modulation of ghrelin receptor in a subject in needthereof, the method comprising administering an effective amount of aGOAT inhibitor. In another embodiment, the invention provides a methodfor the modulation of ghrelin receptor in a subject in need thereof, themethod comprising administering an effective amount of a ghrelinreceptor antagonist and/or inverse agonist.

In another embodiment, the invention provides a method for treatment,prevention, or management of metabolic syndrome in a subject in needthereof, the method comprising the step of administering an effectiveamount of a GOAT inhibitor and an effective amount of a ghrelin receptorantagonist and/or inverse agonist.

In another embodiment, the invention provides a method for treatment,prevention, or management of cancer in a subject in need thereof, themethod comprising the step of administering an effective amount of aGOAT inhibitor and an effective amount of a ghrelin receptor antagonistand/or inverse agonist. In one embodiment, the invention provides amethod for treatment, prevention, or management of cancer in a subjectin need thereof, the method comprising administering an effective amountof a GOAT inhibitor. In another embodiment, the invention provides amethod for treatment, prevention, or management of cancer in a subjectin need thereof, the method comprising administering an effective amountof a ghrelin receptor antagonist and/or inverse agonist.

In one embodiment, the GOAT inhibitor comprises an octanoylated peptideand the octanoylation is at position three of the peptide. In anotherembodiment, the GOAT inhibitor comprises an octanoylated pentapeptideand wherein the octanoylation is at position three of the pentapeptide.In one embodiment, the octanoylated ghrelin pentapeptide isGly-Ser-Ser(Oct)-Phe-Leu (SEQ ID NO: 1), or Gly-Ser-Dap(Oct)-Phe-Leu(SEQ ID NO: 2). Ser(Oct) and Dap(Oct) represent octanoylated serine andoctanoylated diaminopropionic acid respectively.

In one embodiment, the ghrelin receptor antagonist is selected from thegroup consisting of Tyr-DTrp-DLys-Trp-DPhe-NH₂ (SEQ ID NO: 11),Tyr-DTrp-Lys-Trp-DPhe-NH₂ (SEQ ID NO: 12), His-DTrp-DLys-Trp-DPhe-NH₂(SEQ ID NO: 13), His-DTrp-DLys-Phe-DTrp-NH₂ (SEQ ID NO: 14),His-DTrp-DArg-Trp-DPhe-NH₂ (SEQ ID NO: 15),His-DTrp-DLys-Trp-DPhe-Lys-NH₂ (SEQ ID NO: 16),DesaminoTyr-DTrp-Ala-Trp-DPhe-NH₂ (SEQ ID NO: 17),DesaminoTyr-DTrp-DLys-Trp-DPhe-NH₂ (SEQ ID NO: 18),DeaminoTyr-DTrp-Ser-Trp-DPhe-Lys-NH₂ (SEQ ID NO: 19),DesaminoTyr-DTrp-Ser-Trp-DPhe-NH₂ (SEQ ID NO: 20),His-DTrp-DTrp-Phe-Met-NH₂ (SEQ ID NO: 21), Tyr-DTrp-DTrp-Phe-Phe-NH₂(SEQ ID NO: 22), Glyψ[CH₂NH]-DβNal-Ala-Trp-DPhe-Lys-NH₂ (SEQ ID NO: 23),Glyψ[CH₂NH]-DβNal-DLys-Trp-DPhe-Lys-NH₂ (SEQ ID NO: 24),DAla-DβNal-DLys-DTrp-Phe-Lys-NH₂ (SEQ ID NO: 25),His-DβNal-DLys-Trp-DPhe-Lys-NH₂ (SEQ ID NO: 26),Ala-His-DTrp-DLys-Trp-DPhe-Lys-NH₂ (SEQ ID NO: 27),Alaψ[CH₂NH]-DβNal-Ala-Trp-DPhe-Lys-NH₂ (SEQ ID NO: 28),DβNal-Ala-Trp-DPhe-Ala-NH₂ (SEQ ID NO: 29),DAla-DcyclohexylAla-Ala-Phe-DPhe-Nle-NH₂ (SEQ ID NO: 30),DcyclohexylAla-Ala-Phe-DTrp-Lys-NH₂ (SEQ ID NO: 31),DAla-DβNal-Ala-Thr-DThr-Lys-NH₂ (SEQ ID NO: 32),DcyclohexylAla-Ala-Trp-DPhe-NH₂ (SEQ ID NO: 33),DAla-DβNal-Ala-Ala-DAla-Lys-NH₂ (SEQ ID NO: 34),DβNal-Ala-Trp-DPhe-Leu-NH₂ (SEQ ID NO: 35),His-DTrp-Phe-Trp-DPhe-Lys-NH₂ (SEQ ID NO: 36),DAla-DβNal-DAla-DTrp-Phe-Lys-NH₂ (SEQ ID NO: 37),βAla-Trp-DAla-DTrp-Phe-NH₂ (SEQ ID NO: 38), His-Trp-DAla-DTrp-Phe-LysNH₂(SEQ ID NO: 39), DLys-DβNal-Ala-Trp-DPhe-Lys-NH₂ (SEQ ID NO: 40),DAla-DβNal-DLys-DTrp-Phe-Lys-NH₂ (SEQ ID NO: 41), Tyr-DAla-Phe-Aib-NH₂(SEQ ID NO: 42), Tyr-DAla-Sar-NMePhe-NH₂ (SEQ ID NO: 43),αγAbu-DTrp-DTrp-Ser-NH₂ (SEQ ID NO: 44), αγAbu-DTrp-DTrp-Lys-NH₂ (SEQ IDNO: 45), αγAbu-DTrp-DTrp-Orn-NH₂ (SEQ ID NO: 46), αAbu-DTrp-DTrp-Orn-NH₂(SEQ ID NO: 47), DThr-DαNal-DTrp-DPro-Arg-NH₂ (SEQ ID NO: 48),DAla-Ala-DAla-DTrp-Phe-Lys-NH₂ (SEQ ID NO: 49),Alaψ[CH₂NH]His-DTrp-Ala-Trp-DPhe-Lys-NH₂ (SEQ ID NO: 50),Lys-DHis-DTrp-Phe-NH₂ (SEQ ID NO: 51), γAbu-DTrp-DTrp-Orn-NH₂ (SEQ IDNO: 52), inip-Trp-Trp-Phe-NH₂ (SEQ ID NO: 53), Ac-DTrp-Phe-DTrp-Leu-NH₂(SEQ ID NO: 54), Ac-DTrp-Phe-DTrp-Lys-NH₂ (SEQ ID NO: 55),Ac-DTrp-DTrp-Lys-NH₂ (SEQ ID NO: 56), DLys-Tyr-DTrp-DTrp-Phe-Lys-NH₂(SEQ ID NO: 57), Ac-DβNal-Leu-Pro-NH₂ (SEQ ID NO: 58),βAla-Trp-DTrp-DTrp-Orn-NH₂ (SEQ ID NO: 59), DVal-DαNal-DTrp-Phe-Arg-NH₂(SEQ ID NO: 60), DLeu-DαNal-DTrp-Phe-Arg-NH₂ (SEQ ID NO: 61),CyclohexylAla-DαNal-DTrp-Phe-Arg-NH₂ (SEQ ID NO: 62),DTrp-DαNal-DTrp-Phe-Arg-NH₂ (SEQ ID NO: 63), DAla-DβNal-DPro-Phe-Arg-NH₂(SEQ ID NO: 64), Ac-DαNal-DTrp-Phe-Arg-NH₂ (SEQ ID NO: 65),DαNal-DTrp-Phe-Arg-NH₂ (SEQ ID NO: 66), His-DTrp-DTrp-Lys-NH₂ (SEQ IDNO: 67), Ac-DβNal-DTrp-NH₂ (SEQ ID NO: 68), αAib-DTrp-DcyclohexylAla-NH₂(SEQ ID NO: 69), αAib-DTrp-DAla-cyclohexylAla-NH₂ (SEQ ID NO: 70),DAla-DcyclohexylAla-Ala-Ala-Phe-DPhe-Nle-NH₂ (SEQ ID NO: 71),DPhe-Ala-Phe-DPa1-NH₂ (SEQ ID NO: 72), DPhe-Ala-Phe-DPhe-Lys-NH₂ (SEQ IDNO: 73), DLys-Tyr-DTrp-DTrp-Phe-NH₂ (SEQ ID NO: 74),Ac-DLys-Tyr-DTrp-DTrp-Phe-NH₂ (SEQ ID NO: 75),Arg-DTrp-Leu-Tyr-Trp-Pro(cyclic Arg-Pro) (SEQ ID NO: 76),Ac-DβNal-PicLys-ILys-DPhe-NH₂ (SEQ ID NO: 77), DPal-Phe-DTrp-Phe-Met-NH₂(SEQ ID NO: 78), DPhe-Trp-DPhe-Phe-Met-NH₂ (SEQ ID NO: 79),DPal-Trp-DPhe-Phe-Met-NH₂ (SEQ ID NO: 80), βAla-Pal-DTrp-DTrp-Orn-NH₂(SEQ ID NO: 81), αγAbu-Trp-DTrp-DTrp-Orn-NH₂ (SEQ ID NO: 82),βAla-Trp-DTrp-DTrp-Lys-NH₂ (SEQ ID NO: 83), γAbu-Trp-DTrp-DTrp-Orn-NH₂(SEQ ID NO: 84), Ava-Trp-DTrp-DTrp-Orn-NH₂ (SEQ ID NO: 85),DLys-Tyr-DTrp-Ala-Trp-DPhe-NH₂ (SEQ ID NO: 86),His-DTrp-DArg-Trp-DPhe-NH₂ (SEQ ID NO: 87), <Glu-His-Trp-DSer-DArg-NH₂(SEQ ID NO: 88), DPhe-DPhe-DTrp-Met-DLys-NH₂ (SEQ ID NO: 89),Gly-Met-Ala-Gly-Ser-(Dap-Oct)-Phe-Leu-Ser-Pro-Glu-His-NH₂ (SEQ ID NO:3), Gly-Met-Ala-Gly-Ser-(Dap-palmityl)-Phe-Leu-Ser-Pro-Glu-His-NH₂ (SEQID NO: 4), O-(2-methylallyl)benzophenone oxime,(R)-2-amino-3-(1H-indol-3-yl)-1-(4-phenylpiperidin-1-yl)propan-1-one,N—((R)-1-((R)-1-((S)-3-(1H-indol-3-yl)-1-oxo-1-(4-phenylpiperidin-1-yl)propan-2-ylamino)-6-amino-1-oxohexan-2-ylamino)-3-hydroxy-1-oxopropan-2-yl)benzamide,(S)—N—((S)-3-(1H-indol-3-yl)-1-oxo-1-(4-phenylpiperidin-1-yl)propan-2-yl)-6-acetamido-2-((S)-2-amino-3-(benzyloxy)propanamido)hexanamide,(S)—N—((R)-3-(1H-indol-3-yl)-1-oxo-1-(4-phenylpiperidin-1-yl)propan-2-yl)-2-((S)-2-acetamido-3-(benzyloxy)propanamido)-6-aminohexanamide,(R)—N-(3-(1H-indol-3-yl)-1-(4-(2-methoxyphenyl)piperidin-1-yl)-1-oxopropan-2-yl)-4-aminobutanamide,(R)—N-(3-(1H-indol-3-yl)-1-(4-(2-methoxyphenyl)piperidin-1-yl)-1-oxopropan-2-yl)-2-amino-2-methylpropanamide,methyl 3-(p-tolylcarbamoyl)-2-naphthoate, ethyl3-(4-(2-methoxyphenyl)piperidine-1-carbonyl)-2-naphthoate,3-(2-methoxyphenylcarbamoyl)-2-naphthoate,(S)-2,4-diamino-N—((R)-3-(naphthalen-2-ylmethoxy)-1-oxo-1-(4-phenylpiperidin-1-yl)propan-2-yl)butanamide,naphthalene-2,3-diylbis((4-(2-methoxyphenyl)piperazin-1-yl)methanone),(R)-2-amino-N-(3-(benzyloxy)-1-oxo-1-(4-phenylpiperazin-1-yl)propan-2-yl)-2-methylpropanamide,or (R)-2-amino-3-(benzyloxy)-1-(4-phenylpiperazin-1-yl)propan-1-one, andpharmaceutically acceptable salts, prodrugs, or active metabolitesthereof.

In some embodiments, the ghrelin receptor antagonist isGly-Met-Ala-Gly-Ser-(Dap-Oct)-Phe-Leu-Ser-Pro-Glu-His-NH₂ (SEQ ID NO:3).

BRIEF DESCRIPTION OF THE FIGURES

FIGS. 1A and 1B show mean plasma GH responses to 500 ng ghrelinadministered icy 15 min after the icy injection of 5 μg GHS-A (FIG. 1B)or normal saline (FIG. 1A). Central pretreatment with GHS-A abolishedthe stimulatory action of ghrelin on GH release compared with normalsaline i.c.v. pretreated controls. Values are the mean±SE. The number ofanimals in each group is shown in parentheses. Arrows indicate the timesof i.c.v. injections.

FIGS. 2A and 2B show mean plasma GH responses to 5 μg ghrelinadministered iv 15 min after the iv injection of 250 μg GHS-A (FIG. 2B)or normal saline (FIG. 2A). Peripheral administration of GHS-A stronglyblocked ghrelin's ability to release GH compared with normalsaline-pretreated controls. Values are the mean±SE. The number ofanimals in each group is shown in parentheses. Arrows indicate the timesof i.v. injections.

FIG. 3 shows a summary of the effects of GHS-A, given centrally (icy) orperipherally (iv), on ghrelin-induced GH release. The GH AUC followingi.c.v. (500 ng) and i.v. (5 μg) ghrelin injection was reduced by 15- and5-fold, respectively, in the GHS-A pretreated groups compared with theirrespective normal saline-treated controls. Each bar represents themean±SE. *, P<0.0003 or less compared with normal saline-pretreatedanimals.

FIGS. 4A and 4B show that Individual representative plasma GH profilesin rats i.c.v. administered either 5 μg GHS-A (FIG. 4B) or normal saline(FIG. 4A) 15 min prior to the expected onset of the spontaneous GHsecretory bursts typical of the male rat. GHS-A administration severelyattenuated the amplitude of the spontaneous GH pulses compared withnormal saline icy-injected controls. Arrows indicate the times of i.c.v.injections.

FIG. 5 shows that the 1-h GH AUC's of the spontaneous GH secretoryepisodes at 1100 h and 1400 h, and the overall 6-h GH AUC, weresignificantly reduced in animals treated i.c.v. with 5 μg GHS-A comparedwith normal saline i.c.v.-treated controls. Values are the mean±SE. *,P<0.01 or less compared with normal saline i.c.v.-treated group.

FIG. 6 shows a cluster analysis of the effects of centrally-administeredGHS-A (5 μg) or normal saline on spontaneous GH pulse parameters.Cluster analysis revealed a significant suppression of GH peak height,but no significant effect of GHS-A on any other parameters of GHpulsatility, including GH peak frequency, interpeak interval and nadir,compared with normal saline icy-treated controls. Values are themean±SE. *, P<0.03 vs. normal saline i.c.v.-treated controls.

FIGS. 7A and 7B show a feeding response to icy-administered ghrelin (500ng) in animals pretreated icy with either GHS-A (5 μg) or normal saline(FIG. 7A). GHS-A significantly inhibited ghrelin's stimulatory effectson food intake in the first hour after injections, compared with normalsaline i.c.v.-pretreated controls (FIG. 7B). Cumulative food intake wassignificantly suppressed for up to 5 h after GHS-A injection. Values arethe mean±SE. *, P<0.02 or less compared with normal salineicv-pretreated controls.

FIGS. 8A and 8B show the effects of icv-administered GHS-A (5 μg) ornormal saline on spontaneous food intake in overnight-fasted animals(FIG. 8A). GHS-A significantly inhibited spontaneous food intake in thefirst hour after injection, compared with normal saline icy-treatedcontrols (FIG. 8B). Cumulative food intake was not inhibited by GHS-Abeyond the first hour after injection. Values are the mean±SE. *,P<0.004 compared with normal saline icy-treated controls.

FIG. 9 shows a diagram of one ghrelin receptor antagonistHisDβNalDLysTrpDPheLysNH₂ (SEQ ID NO: 26).

FIG. 10 shows GMAGS(Dap-Oct)FLSPEH-NH₂ (peptide #3) (SEQ ID NO: 3), butnot GMAGS(Dap-Palmityl)FLSPEH-NH₂ (peptide #6) (SEQ ID NO: 4) inhibitedfood intake. FIG. 10 discloses “GMAGS” as residues 1-5 of SEQ ID NOS 3and 4.

FIG. 11 shows GMAGS(Dap-Oct)FLSPEH-NH₂ (peptide #3) (SEQ ID NO: 3), butnot GMAGS(Dap-Palmityl)FLSPEH-NH₂ (peptide #6) (SEQ ID NO: 4) inhibitedgrowth hormone secretion. FIG. 11 discloses “GMAGS” as residues 1-5 ofSEQ ID NOS 3 and 4.

DETAILED DESCRIPTION OF THE INVENTION

The present invention provides novel strategies for inhibiting and/ordisrupting the GHS-R 1a signaling pathway for regulating GH secretion,appetite, and body weight, and for the treatment of metabolic diseasesand disorders such as obesity, overeating, diabetes mellitus,unregulated cell proliferation, and for the inhibition of growth hormonesecreted from tumors such as pituitary, prostate, medullary thyroidcarcinomas, osteoblast, pancreatic and hepatoma. The inhibition and/ordisruption of the GHS-R signaling pathway can be approached in severalways: (1) inhibiting of synthesis of the active form of ghrelin, anendogenous peptide ligand of the pathway and (2) disrupting theconstitutively active GHS-R 1a signaling pathway using antagonistsand/or inverse agonists. These approaches can be applied individually ortogether.

Ghrelin, a 28 amino acid, octanoylated, appetite-stimulating peptidehormone, is secreted by the food-deprived stomach. It is the endogenousligand for GHS-R 1a and therefore, the activator ligand of the GHS-R 1asignaling pathway. Ghrelin is synthesized as a preprohormone, thenproteolytically processed to yield a 28-amino acid peptide. Aninteresting and unique modification is then imposed on the 28-amino acidpeptide hormone during synthesis. This peptide hormone requiresacylation with an eight-carbon fatty acid, octanoate, at amino acidresidue serine-3 (Bednarek M A. et al., 2000, J Med Chem., 43:4370-6;Kojima et al., 1999, Nature, 402:656-60, content of both of which isherein incorporated by reference). This modification is necessary forbiologic activity, i. e. activation of intracellular signaling that ismediated through the seven transmembrane G (7TMG) protein coupledghrelin receptor (GHS-R 1a)—both in vitro and in vivo (Kojima et al.,1999, Nature, 402:656-60; Nakazato et al., 2001, Nature, 409:194-8;Tschop et al., 2000, Nature, 407:908-13, content of all of which isherein incorporated by reference). Ghrelin that is deleted of theoctanoate, known as desoctanoylated ghrelin, is biologically inactive.

Recently, the membrane bound acyltransferase that catalyses the additionof octanoate to serine-3 was identified. The enzyme is named ghrelinO-acyltransferase (GOAT). GOAT was shown to also transfer octanoyl to apentapeptide containing only the N-terminal five amino acids ofproghrelin, the 94 amino acid protein precursor of ghrelin (Yang J, et.al., 2008, Proc. Natl. Acad. Sci. USA. 105:10750-5, content of which isherein incorporated by reference). Yang, et, al. showed that GOATactivity could be inhibited by an octanoylated ghrelin pentapeptide, andits potency was enhanced 45-fold when the octanoylated serine-3 wasreplaced by octanoylated diaminopropionic acid.

Accordingly, strategies for inhibiting the GHS-R 1a signaling pathwayencompass preventing the synthesis of an active form of ghrelin viainhibiting the activity of GOAT with an octanoylated ghrelinpentapeptide or octanoylated diaminopropionic peptides and the likes.

Synthesis of ghrelin occurs predominantly in P/D1 epithelial cellslining the fundus of the stomach and epsilon cells of the pancreas thatstimulates appetite. Ghrelin levels increase before meals and decreaseafter meals. Smaller amounts of ghrelin is produced in the placenta,kidney, pituitary and hypothalamus. In addition, certain tumors andcancers have been shown to express ghrelin although the normal tissue donot: islet cell tumors, medullary thyroid carcinomas, pituitary adenoma,thyroid tumor, and pancreatic and gastrointestinal endocrine tumors(Korbonits M, et. al. 2001, J. Clin. Endocrinol. Metab. 86:881-887;Papotti M, et. al., 2001, J. Clin. Endocrinol. Metab. 86:5052-5059;Kanamoto N, et. al., 2001, J. Clin. Endocrinol. Metab. 86:4984-4990;Korbonits M, et. al., 2001, Endocrine 14:101-104; Volante M, et. al.2002, J. Clin. Endocrinol. Metab. 87:1300-1308, content of all of whichis herein incorporated by reference).

The ghrelin receptor, GHS-R 1a, is found in cells within the anteriorpituitary which when activated, potently stimulates secretion of growthhormone. Ghrelin receptors are present on the cells in the pituitarythat secrete growth hormone, and also have been identified in thehypothalamus, heart and adipose tissue.

Interaction of the active octanoylated peptide hormone with itsreceptor, the ghrelin receptor, GHS-R 1a, leads to the release of growthhormone and the positive activation of the GH axis. The ghrelin/GHS-R 1asignaling pathway is also involved in the regulation of energy balancein the body. Regulation of energy balance comprises ghrelin functions toincrease hunger though its action on hypothalamic feeding centers,suppress fat utilization in adipose tissue, stimulating gastric emptyingand having a variety of positive effects on cardiovascular function(e.g. increased cardiac output). The ultimate effect of ghrelin is thestimulation of appetite, intake of food and the secretion of growthhormone. Thus, any strategies for inhibiting and/or disrupting theghrelin/GHS-R 1a signaling pathway which includes reducing the amount ofcirculating ghrelin can be more effective in the treatment of metabolicdiseases and disorders associated with abnormal appetite and intake offood, energy balance and regulation, and/or ectopic release of growthhormone and/or ghrelin such as induction by tumors.

In one embodiment, the method for inhibiting and/or disrupting theghrelin/GHS-R 1a signaling pathway comprises inhibiting the synthesis ofan active octanoylated ghrelin comprising inhibiting a ghrelinO-acyltransferase (GOAT).

In one embodiment, the method for inhibiting and/or disrupting theghrelin/GHS-R 1a signaling pathway comprises inhibiting the constitutiveghrelin/GHS-R 1a signaling pathway with an inverse agonist.

In one embodiment, the method for inhibiting and/or disrupting theghrelin/GHS-R 1a signaling pathway comprises inhibiting the constitutiveghrelin/GHS-R 1a signaling pathway comprising inhibiting the interactionof GHS-R 1a and its ligand, e. g. ghrelin, with an antagonist.

In one embodiment, the method for inhibiting and/or disrupting theghrelin/GHS-R 1a signaling pathway comprises simultaneously inhibitingthe synthesis of active octanoylated ghrelin and inhibiting theinteraction of the active octanoylated hormone with its receptor, theghrelin receptor, GHS-R 1a and/or the constitutive GHS-R 1a signalingpathway. For example, by administering an effective amount of a ghrelinO-acyltransferase (GOAT) inhibitor and an effective amount of a ghrelinreceptor antagonist and/or inverse agonist. While not wishing to bebound by theory, inhibiting the synthesis of active octanoylated ghrelinserves to reduce the amount of circulating ghrelin that is available forbinding to and activating the ghrelin receptor, GHS-R 1a, and theassociated signaling pathway Inhibiting the interaction of the activeoctanoylated ghrelin with GHS-R 1a serves to inhibit the ghrelin/GHS-R1a signaling pathway. The inverse agonist serves to reduce the amount ofconstitutive signaling from the pathway. The combined inhibitionstrategy work synergistically to attenuates GH pulses and reduces foodintake in mammals.

The serine-3 of ghrelin is acylated with an eight-carbon fatty acid,octanoate, which is required for its endocrine actions. Ghrelin that isdeleted of the octanoate, known as desoctanoylated ghrelin isbiologically inactive. The membrane bound acyltransferase that catalysesthe addition of octanoate to Serine-3 (Yang, et, al. 2008,) namedghrelin O-acyltransferase (GOAT). GOAT was shown to also transferoctanoyl to a pentapeptide containing only the N-terminal five aminoacids of proghrelin, the 94 amino acid protein precursor of ghrelin(Yang, et, al. 2008). Yang, et, al. 2008 showed that GOAT activity couldbe inhibited by an octanoylated ghrelin pentapeptide, and its potencywas enhanced 45-fold when the octanoylated serine-3 was replaced byoctanoylated diaminopropionic acid. Accordingly, the synthesis of activeoctanoylated ghrelin can be inhibited with an octanoylated ghrelinpentapeptide or modified versions thereof, such as an octanoylateddiaminopropionic peptide.

The interaction of the active octanoylated hormone with its receptor,the ghrelin receptor, GHS-R 1a, can be inhibited with a ghrelin receptorantagonist and/or a growth hormone secretatogue antagonist and/orinverse agonist.

In one embodiment, provided herein is a method of treatment, preventionor management of obesity in a subject, the method comprisingadministering an effective amount of a ghrelin O-acyltransferase (GOAT)inhibitor and an effective amount of a ghrelin receptor antagonistand/or a growth hormone secretatogue antagonist and/or inverse agonist.

In one embodiment, provided herein is a method of treatment, preventionor management of diabetes mellitus in a subject, the method comprisingadministering an effective amount of a ghrelin O-acyltransferase (GOAT)inhibitor and an effective amount of a ghrelin receptor antagonistand/or a growth hormone secretatogue antagonist and/or inverse agonist.

In one embodiment, provided herein is a method of treatment, preventionor management of metabolic syndrome in a subject, the method comprisingadministering an effective amount of a ghrelin O-acyltransferase (GOAT)inhibitor and an effective amount of a ghrelin receptor antagonistand/or a growth hormone secretatogue antagonist and/or inverse agonist.

In one embodiment, provided herein is a method of treatment, preventionor management of cancer in a subject, the method comprisingadministering an effective amount of a ghrelin O-acyltransferase (GOAT)inhibitor and an effective amount of a ghrelin receptor antagonistand/or a growth hormone secretatogue antagonist and/or inverse agonist.In one embodiment, the method for treatment, prevention, or managementof cancer in a subject in need thereof comprises administering aneffective amount of a GOAT inhibitor. In another embodiment, the methodfor treatment, prevention, or management of cancer in a subject in needthereof comprises administering an effective amount of a ghrelinreceptor antagonist and/or inverse agonist and/or a growth hormonesecretatogue antagonist.

In some embodiments, the methods of treatment, prevention or managementof obesity, diabetes mellitus, metabolic syndrome, or cancer in asubject in need hereof comprise administering an effective amount of aghrelin O-acyltransferase (GOAT) inhibitor.

In other embodiments, the methods of treatment, prevention or managementof obesity, diabetes mellitus, metabolic syndrome, or cancer in asubject in need hereof comprise administering an effective amount of aghrelin receptor antagonist and/or a growth hormone secretatogueantagonist and/or inverse agonist.

In one embodiment, provided herein is a method of modulating a ghrelinreceptor in a subject, the method comprising administering an effectiveamount of a ghrelin O-acyltransferase (GOAT) inhibitor and an effectiveamount of a ghrelin receptor antagonist and/or a growth hormonesecretatogue antagonist. In another embodiment, provided herein is amethod of modulating a ghrelin receptor in a subject, the methodcomprising administering an effective amount of a ghrelinO-acyltransferase (GOAT) inhibitor. In yet another embodiment, providedherein is a method of modulating a ghrelin receptor in a subject, themethod comprising administering an effective amount of a ghrelinreceptor antagonist and/or inverse agonist.

As used herein, the term “modulation” specifically refers to theinhibition of ghrelin receptor activity such as ghrelin ligand binding,message transduction, increased intracellular inositol1,4,5-trisphosphate (IP3) levels and any activity associated with theactivation and generation of intracellular signals in theghrelin/ghrelin receptor signaling pathway. Methods of receptor-ligandbinding and determining levels of IP3 are well known in the art, e.g.,as described by AR Prasad, et al., 1993, Circulation Research,72:827-836, content of which is herein incorporated by reference.

In one embodiment, provided herein is a method of reducing a desire of ahuman subject to consume calories following gastric banding or gastricbypass surgery, the method comprising administering an effective amountof a ghrelin O-acyltransferase (GOAT) inhibitor and an effective amountof a ghrelin receptor antagonist and/or a growth hormone secretatogueantagonist and/or inverse agonist. In another embodiment, the method ofreducing a desire of a human subject to consume calories followinggastric banding or gastric bypass surgery comprises administering aneffective amount of a ghrelin O-acyltransferase (GOAT) inhibitor. Inanother embodiment, the method of reducing a desire of a human subjectto consume calories following gastric banding or gastric bypass surgerycomprises administering an effective amount of a ghrelin receptorantagonist and/or a growth hormone secretatogue antagonist and/orinverse agonist.

In one embodiment, provided herein is a method of treating hormonallyfunctional endocrine or non-endocrine tumors in a subject, the methodcomprising administering an effective amount of a ghrelinO-acyltransferase (GOAT) inhibitor and an effective amount of a ghrelinreceptor antagonist and/or a growth hormone secretatogue antagonistand/or inverse agonist. Such as, but not limited to, pituitary tumors,including ACTH-secreting pituitary tumors, in a mammal. In anotherembodiment, the method of treating hormonally functional endocrine ornon-endocrine tumors in a subject comprises administering an effectiveamount of a ghrelin O-acyltransferase (GOAT) inhibitor. In anotherembodiment, the method of treating hormonally functional endocrine ornon-endocrine tumors in a subject comprises administering an effectiveamount of a ghrelin receptor antagonist and/or a growth hormonesecretatogue antagonist and/or inverse agonist. The inventive methodsinvolve administering to the mammal having or at risk for developing apituitary tumor a therapeutically effective amount of a ghrelinO-acyltransferase (GOAT) inhibitor and/or an effective amount of aghrelin receptor antagonist and/or a growth hormone secretatogueantagonist. In one embodiment, the ghrelin receptor antagonist and/orGOAT inhibitor are administered in combination with other compoundsuseful in the treatment of pituitary tumor such as, for example, PPARγligands. PPARγ ligands include thiazolidinediones (TZDs), such astroglitazone, pioglitazone, and rosiglitazone.

In one embodiment, provided herein is a method for the treatment oftumors that produce prolactin in a subject, the method comprisingadministering an effective amount of a ghrelin O-acyltransferase (GOAT)inhibitor and an effective amount of a ghrelin receptor antagonistand/or a growth hormone secretatogue antagonist and/or inverse agonist.In another embodiment, the method for the treatment of tumors thatproduce prolactin in a subject comprises administering an effectiveamount of a ghrelin O-acyltransferase (GOAT) inhibitor. In anotherembodiment, the method for the treatment of tumors that produceprolactin in a subject comprises administering an effective amount of aghrelin receptor antagonist and/or a growth hormone secretatogueantagonist and/or inverse agonist. Such tumors include, but are notlimited to, breast, pituitary, and prostate cancer. A therapeuticallyeffective amount of a GOAT inhibitor and/or an effective amount of aghrelin receptor antagonist and/or a growth hormone secretatogueantagonist are administered to a subject with or at risk for developinga tumor that produces prolactin.

In one embodiment, provided herein is a method of inhibitingadrenacorticotropic hormone in a subject in need thereof, the methodcomprising administering an effective amount of a ghrelinO-acyltransferase (GOAT) inhibitor and an effective amount of a ghrelinreceptor antagonist and/or a growth hormone secretatogue antagonistand/or inverse agonist. In another embodiment, the method of inhibitingadrenacorticotropic hormone in a subject in need thereof comprisesadministering an effective amount of a ghrelin O-acyltransferase (GOAT)inhibitor. In another embodiment, the method of inhibitingadrenacorticotropic hormone in a subject in need thereof comprisesadministering an effective amount of a ghrelin receptor antagonistand/or a growth hormone secretatogue antagonist and/or inverse agonist.A subject in need of inhibiting adrenacorticotropic hormone is one withtoo much adrenacorticotropic hormone. Excessive adrenacorticotropichormone (ACTH) results in Cushing's disease. Physical symptoms of thedisease include but are not limited to widened face with acne andflushing, fatty deposits over back of neck, stretch marks, easybruising, hair growth, diabetes mellitus, muscle loss and fatigue, anddepression and psychosis. A routine blood test that is well known to oneskilled in the art can be used to determine the level of the hormone inthe individual. Adrenacorticotropic hormone are produced byneuroendocrine tumors, carcinoid, pituitary and pancreatic tumors.Accordingly, in another embodiment, provided herein is a method oftreating ectopic neuroendocrine tumors, the method comprisingadministering an effective amount of a ghrelin O-acyltransferase (GOAT)inhibitor, and/or an effective amount of a ghrelin receptor antagonistand/or a growth hormone secretatogue antagonist and/or inverse agonist.

In one embodiment, provided herein is a method of inhibiting signalingvia subtype receptor of ghrelin in addition to GHS-type 1 receptor in asubject in need thereof, the method comprising administering aneffective amount of a ghrelin O-acyltransferase (GOAT) inhibitor and aneffective amount of a ghrelin receptor antagonist and/or a growthhormone secretatogue antagonist and/or inverse agonist. In anotherembodiment, the method of inhibiting signaling via subtype receptor ofghrelin in addition to GHS-type 1 receptor in a subject in need thereofcomprises administering an effective amount of a ghrelinO-acyltransferase (GOAT) inhibitor. In another embodiment, the method ofinhibiting signaling via subtype receptor of ghrelin in addition toGHS-type 1 receptor in a subject in need thereof comprises administeringan effective amount of a ghrelin receptor antagonist and/or a growthhormone secretatogue antagonist and/or inverse agonist. Skilled artisanis well aware of diseases and disorders associated with these subtypereceptors, e.g. subtype 1b receptor. For example, a review of thesubtype receptors is found in J. P. Camilla, 2005, J. Neuroendocrin.,18: 65-76, content of which is herein incorporated by reference. Certaincancers such as prostate cancer, osteoblast cancer, pancreatic cancer,adenocarcinomas and hepatoma cells are associated with different subtypeof GHS-type 1 receptor. Accordingly, in another embodiment, providedherein is a method of treating diseases and disorders associated withghrelin subtype receptors in a subject in need thereof, the methodcomprising administering an effective amount of a ghrelinO-acyltransferase (GOAT) inhibitor and an effective amount of a ghrelinreceptor antagonist and/or a growth hormone secretatogue antagonistand/or inverse agonist. In one embodiment, the method of treatingdiseases and disorders associated with ghrelin subtype receptors in asubject in need thereof comprises administering an effective amount of aghrelin O-acyltransferase (GOAT) inhibitor. In one embodiment, themethod of treating diseases and disorders associated with ghrelinsubtype receptors in a subject in need thereof comprises administeringan effective amount of a ghrelin receptor antagonist and/or a growthhormone secretatogue antagonist and/or inverse agonist.

In one embodiment, provided herein is a method of treatment, preventionor management of obesity related diseases and disorders in a subject,the method comprising administering an effective amount of a ghrelinO-acyltransferase (GOAT) inhibitor and an effective amount of a ghrelinreceptor antagonist and/or a growth hormone secretatogue antagonistand/or inverse agonist. In one embodiment, the method of treatment,prevention or management of obesity related diseases and disorders in asubject comprises administering an effective amount of a ghrelinO-acyltransferase (GOAT) inhibitor. In another embodiment, the method oftreatment, prevention or management of obesity related diseases anddisorders in a subject comprises administering an effective amount of aghrelin receptor antagonist and/or a growth hormone secretatogueantagonist and/or inverse agonist. Obesity related diseases anddisorders include, but not limited to overeating, diabetes mellitus,metabolic syndrome, hypertension, elevated plasma insulinconcentrations, insulin resistance, dyslipidemias, and hyperlipidemia.

In some embodiments, the methods of treatment, prevention or managementof obesity, or obesity related diseases and disorders in a subjectfurther comprises administering an anti-obesity treatment. In someembodiment, the anti-obesity treatment is a combination of dietaryrestriction therapy with a 5HT (serotonin) transporter inhibitor, a NE(norepinephrine) transporter inhibitor, a CB-1 (cannabinoid-1)antagonist/inverse agonist, a H3 (histamine H3) antagonist/inverseagonist, a MCH1R (melanin concentrating hormone 1R) antagonist, a MCH2Ragonist/antagonist, a NPY1 antagonist, a leptin, a leptin derivative, aleptin analog, PYY(1-36), PYY(3-36), an opioid antagonist, an orexinantagonist, a BRS3 (bombesin receptor subtype 3) agonist, a CCK-A(cholecystokinin-A) agonist, a CNTF (Ciliary neurotrophic factor), aCNTF derivative, a lipase drug inhibitor, an inhibitor of food intake,an incretin, an incretin agonist, an incretin analog or an incretinmimic administered simultaneously, concurrently or sequentially.

Examples of the inhibitor of food intake include but are not limited togastrointestinal hormone glucagon-like-peptide (Glip-1/Glip-2) and theintestinal preproghrelin derived peptide hormone oxyntomodulin and theiranalogs, derivatives, mimics. Examples of incretins and its agonists,analogs, derivatives, or mimics capable of inducing a decrease in foodintake, include but are not limited to GLP-1 receptor agonists exenatide(synthetic mimetic of exendin-4), liraglutide, or enzyme glucagon-likepeptide-1(GLP-1) inhibitors of the dipeptidyl peptidase DPP-4 type,i.e., sitagliptin, vildagliptin, saxagliptin, which slow degradation ofGLP-1 and prolong the actions of GLP-1.

The incretin hormones GLP-1 (glucagon-like peptide-1) and GIP areproduced by the endocrine cells of the intestine following ingestion offood. GLP-1 and GIP stimulate insulin secretion from the beta cells ofthe islets of Langerhans in the pancreas. Only GLP-1 causes insulinsecretion in the diabetic state. Exenatide, a synthetic version ofexendin-4, a hormone found in the saliva of the Gila monster, bears a50% amino acid homology to GLP-1 and displays biological propertiessimilar to human glucagon-like peptide-1 (GLP-1). It has a longerhalf-life in vivo. exenatide enhances glucose-dependent insulinsecretion by the pancreatic beta-cell, suppresses inappropriatelyelevated glucagon secretion, and slows gastric emptying.

Accordingly, any treatment intervention that increases the GLP-1 and/orGIP stimulated activities in vivo is contemplated, e. g. GLP-1 analogs,derivatives, mimics and also inhibitors to slow GLP-1 degradation andprolong the actions of GLP-1.

Leptin is a 16 kDa protein hormone that plays a key role in regulatingenergy intake and energy expenditure, including appetite and metabolism.Leptin is produced in white adipose tissue—the major source—brownadipose tissue, placenta (syncytiotrophoblasts), ovaries, skeletalmuscle, stomach (lower part of fundic glands), mammary epithelial cells,bone marrow, pituitary and liver. Leptin circulates at levelsproportional to body fat. It enters the central nervous system (CNS) inproportion to its plasma concentration. Its receptors are found in brainneurons involved in regulating energy intake and expenditure. Leptinsignals to the brain that the body has had enough to eat, or satiety,thus reducing appetite.

Leptin can be recombinantly produced as described in e.g., WO 96/05309;U.S. Pat. No. 5,552,522; U.S. Pat. No. 5,552,523; and U.S. Pat. No.5,552,524, the content of all of which is herein incorporated byreference. Biologically active derivatives or analogs of leptin can alsobe referred as as leptin peptide mimetics. These mimetics can bedesigned and produced by techniques known to those of skill in the art.(See e.g., U.S. Pat. Nos. 4,612,132; 5,643,873, 5,654,276 and 5,866,547,content of all of which is herein incorporated by reference).

Peptide YY (PYY) is a short (36 amino acid) protein released by cells inthe ileum and colon in response to feeding. A small amount of PYY about1-10 percent in esophagus, stomach, duodenum and jejunum. It is alsoknown as PYY, peptide tyrosine tyrosine, or pancreatic peptide YY(3-36).There are two major forms of Peptide YY: PYY(1-36) and PYY(3-36). Inhumans it reduce appetites. PYY concentration in the circulationincreases postprandially (after food ingestion) and decreases byfasting.

In other embodiments, the methods of treatment, prevention or managementof obesity, or obesity related diseases and disorders in a subjectfurther comprises a surgical or mechanical procedure used for thetreatment of obesity. Such procedures include but not limited tobariactric surgeries such as gastric stapling or gastroplasty, andgastric bypass.

In some embodiments of the treatment, prevention or management ofdiabetes mellitus or metabolic syndrome described herein, the ghrelinsecretory receptor inhibitors, e. g. ghrelin receptor antagonist orinverse agonist can be administered alone, sequentially, orconcomitantly with a biguanide (Metformin), a peroxisome proliferatoractivator-receptor alpha (PPAR-alpha) ligand or PPAR-gamma ligand suchas pioglitazone or rosiglitazone which increase circulating adiponectinlevels and/or regulate/augment phosphorylation of adenosinemonophosphate protein kinase (AMP-kinase), a key intracellular regulatorof energy balance, and agents that lower peripheral circulating glucoselevels primarily but not exclusively in Type 2 patients with diabetesmellitus. Biguanides are well known oral antihyperglycemic drugs usedfor diabetes mellitus or prediabetes treatment.

In some embodiments, the ghrelin O-acyltransferase (GOAT) inhibitor inany of the methods described herein comprises an octanoylated ghrelinpentapeptide, and wherein the octanoylation is at position three of thepentapeptide. Preferably, the octanoylation is at the side chain of theresidue at position three. In some embodiments, the octanoylated ghrelinpentapeptide is Gly-Ser-Ser(Oct)-Phe-Leu (SEQ ID NO: 1).

In some embodiments, the octanoylated ghrelin pentapeptide isGly-Ser-Dap(Oct)-Phe-Leu (SEQ ID NO: 2).

Other GOAT inhibitors amenable to the present invention are described,for example, in U.S. Pat. Pub. No.: 2010/0086955 and Int. Pat. Pub. No.:WO2010/039461, content of both of which is herein incorporated byreference.

In some embodiment, a mixture of GOAT inhibitors is used in the methodsdescribed herein.

In some embodiments, the ghrelin receptor antagonist is of formula (I):A₁-A₂-A₃-A₄-A₅-A₆, wherein: A₁ is His, Tyr, desamino-Tyr, D- or L-Ala,β-Ala, CyclohexylAla (Cyclohexylalanine), D-Arg, Ava (aminovalericacid), Gly, <Glu (pyroglutaminic acid), α-Aib (alpha-aminoisobutyricacid), γ-Abu (gamma-aminobutyric acid), α-Abu, α,γ-Abu, D-Val, D-phe,D-Thr, D-Pal (pyridylalanine), D-Lys, AcD-Lys, D-Leu, D-Trp,D,α-naphthylalanine, D,β-naphthylalanine, or Ac-D,β-naphthylalanine; A₂is D-α-naphthylalanine, D-β-naphthylalanine; Ac-D-β-naphthylalanine, D-or L-Trp, D- or L-Phe, Ala, His, PicLys (N^(ε)-picoloyl-lysine),D-Cyclohexylalanine, or an amino acid that is methylated at the terminalnitrogen of the a carbon atom of the A₂ residue; A₃ is D- or L-Lys,lysine analogs and derivatives, Arg, arginine analogs and derivatives,Orn, Phe, Trp, Leu, Pro, Ala, Ser, Pal, or α,γ-Abu; A₄ is D- or L-Trp,D- or L-Phe, Ala, Ser, Tyr, Met, Pro, Thr, ILys, or CyclohexylAla; A₅ isD- or L-Trp, D- or L-Phe, Ala, Lys, Arg, Orn, Thr, Leu, orD-CyclohexylAla; A₆ is Lys, Arg, Orn, D- or L-Phe, Pro(cyclic Arg-Pro),Nle (norleucine), α,γ-Abu amide, or a free acid carboxyl group; andprodrugs, metabolite, or pharmaceutically acceptable salts thereof.

In some embodiments of compounds of formula (I), there can be a normalor reduced psi peptide linkage at position 1. Reduced psi-peptidelinkage refers to the peptide linkage (—C(O)NH—) that has been replaceby —CH₂NH—; therefore, reduced psi peptide linkage at position 1 meansthe —C(O)NH— bond between first and second amino acid is replaced by—CH₂NH—. A reduced psi peptide linkage is indicated as ψ[CH₂NH] herein.

In some embodiments, the A₁ is methylated at the terminal nitrogen ofthe alpha carbon atom of the A₁ residue.

In some embodiments, the A₂ has an extended aromatic chain. Exemplary A₂with extended aromatic chains include, but are not limited to,D-4-halo-Phe, D-4-pyrolidylalanine, and homologues or analogues thereof.

In some embodiments, the ghrelin receptor antagonist of formula (I) isHis-DβNal-DLys-Trp-DPhe-Lys-NH₂ (SEQ ID NO: 26) or analogues, prodrug,metabolite, or pharmaceutical salts thereof. In another embodiment, theghrelin receptor antagonist of formula (I) excludesHis-DβNal-DLys-Trp-DPhe-Lys-NH₂ (SEQ ID NO: 26).

In another embodiment, the ghrelin receptor antagonist of formula (I) isHis-DTrp-DLys-Trp-DPhe-Lys-NH₂ (SEQ ID NO: 16)

In some embodiments, the ghrelin receptor antagonist is of formula (II):A₇-A₈-A₉-A₁₀, wherein: A₇ is D-α-Nal, D-β-Nal, Ac-D-β-Nal, Ac-D-α-Nal,D- or L-Tyr, Ac-D-Tyr, Lys, D-Phe, His, α-Abu, α,γ-Abu, γ-Abu,DcyclohexylAla, or isonipecotic carboxylic acid (inip); A₈ is D- orL-Trp, Ala, His, Phe, or Leu; A₉ is D- or L-Trp, Ala, CyclohexylAla,Phe, Pro, Lys, Sarcosine (Sar, N-methylglycine), or a free acid carboxylgroup; A₁₀ is D- or L-Arg, Phe, CyclohexylAla, Lys, Ser, NMePhe, DPal,Aib, or Orn; and prodrugs, metabolite, or pharmaceutically acceptablesalts thereof.

In some embodiments, the ghrelin receptor antagonist of formula (II) isα,γAbu-DTrp-DTrp-Ser-NH₂ (SEQ ID NO: 44), α,γAbu-DTrp-DTrp-Lys-NH₂ (SEQID NO: 45), α,γAbu-DTrp-DTrp-Orn-NH₂ (SEQ ID NO: 46),Tyr-DAla-Phe-Aib-NH₂ (SEQ ID NO: 42), Tyr-DAla-Sar-NMePhe-NH₂ (SEQ IDNO: 43), Lys-DHis-DTrp-Phe-NH₂ (SEQ ID NO: 51), γAbu-DTrp-DTrp-Orn-NH₂(SEQ ID NO: 52), inip-Trp-Trp-Phe-NH₂ (SEQ ID NO: 53),Ac-DTrp-Phe-DTrp-Leu-NH₂ (SEQ ID NO: 54), Ac-DTrp-DTrp-Lys-NH₂ (SEQ IDNO: 56), or Ac-DβNal-Leu-Pro-NH₂ (SEQ ID NO: 58).

In some other embodiments, the ghrelin receptor antagonist of formula(II) is not α,γAbu-DTrp-DTrp-Ser-NH₂ (SEQ ID NO: 44);α,γAbu-DTrp-DTrp-Lys-NH (SEQ ID NO: 45), or α,γAbu-DTrp-DTrp-Orn-NH₂(SEQ ID NO: 46).

In some embodiments, the ghrelin receptor antagonist is of formula(III):

wherein: R₁ is OH; R₂ is H, —C(O)—(CH₂)₃—NH₂, —C(O)—C(CH₃)₂—NH₂,—C(O)—CH((CH₂)₄—NHR₅)—NH—C(O)—CH(NHR₃)CH₂OR₄; R₃ is H, —C(O)-phenyl, or—C(O)CH₃; R₄ is H, or —CH₂-phenyl; R₅ is H, or —C(O)CH₃; and isomers,derivatives, prodrugs, metabolite, or pharmaceutically acceptable saltsthereof.

Exemplary compounds of formula (III) include, but are not limited to,(R)-2-amino-3-(1H-indol-3-yl)-1-(4-phenylpiperidin-1-yl)propan-1-one;N—((R)-1-((R)-1-((S)-3-(1H-indol-3-yl)-1-oxo-1-(4-phenylpiperidin-1-yl)propan-2-ylamino)-6-amino-1-oxohexan-2-ylamino)-3-hydroxy-1-oxopropan-2-yl)-benzamide;(S)—N—((S)-3-(1H-indol-3-yl)-1-oxo-1-(4-phenylpiperidin-1-yl)propan-2-yl)-6-acetamido-2-((S)-2-amino-3-(benzyloxy)propanamido)-hexanamide;(S)—N—((R)-3-(1H-indol-3-yl)-1-oxo-1-(4-phenylpiperidin-1-yl)propan-2-yl)-2-((S)-2-acetamido-3-(benzyloxy)propanamido)-6-aminohexanamide;(R)—N-(3-(1H-indol-3-yl)-1-(4-(2-methoxyphenyl)piperidin-1-yl)-1-oxopropan-2-yl)-4-aminobutanamide;(R)-2-amino-N-(3-(benzyloxy)-1-oxo-1-(4-phenylpiperazin-1-yl)propan-2-yl)-2-methylpropanamide;and isomers and derivatives thereof

In some embodiments, the ghrelin receptor antagonist is of formula (IV):

wherein: R₆ is —C(O)OCH₂CH₃, —C(O)OCH₃,

R₇ is —H,

and isomers, derivatives, prodrugs, metabolite, or pharmaceuticallyacceptable salts thereof.

Exemplary compounds of formula (IV) include, but are not limited to,naphthalene-2,3-diylbis((4-(2-methoxyphenyl)piperazin-1-yl)-methanone);3-(2-methoxyphenylcarbamoyl)-2-naphthoate;(S)-2,4-diamino-N—((R)-3-(naphthalen-2-ylmethoxy)-1-oxo-1-(4-phenylpiperidin-1-yl)propan-2-yl)-butanamide;ethyl 3-(4-(2-methoxyphenyl)-piperidine-1-carbonyl)-2-naphthoate; methyl3-(p-tolylcarbamoyl)-2-naphthoate; and isomers and derivatives thereof.

In some embodiments, the ghrelin receptor antagonist is of formula (V):

wherein: R₈ is H, or —C(O)C(CH₃)₂—NH₂; and isomers, derivatives,prodrugs, metabolite, or pharmaceutically acceptable salts thereof.

Exemplary compounds of formula (V) include, but are not limited to,(R)-2-amino-N-(3-(benzyloxy)-1-oxo-1-(4-phenylpiperazin-1-yl)propan-2-yl)-2-methylpropanamide;(R)-2-amino-3-(benzyloxy)-1-(4-phenylpiperazin-1-yl)propan-1-one; andisomers and derivatives thereof.

In some embodiments, the ghrelin receptor antagonist is of formula (VI):S¹B¹S²B²S³, wherein: S¹ is H, CO₂H, R¹¹, R¹¹R¹², or R¹¹R¹²R¹³; S² isdes-Amino, H, CO₂H, R¹¹, R¹¹R¹², or R¹¹R¹²R¹³; S³ is H, CO₂H, NH₂, R¹¹,R¹¹R¹², or R¹¹R¹²R¹³; R¹¹, R¹², and R¹³ are independently selected fromthe group consisting of L-amino acids, Pal (3-pyridyl alanine),cyclo-Ala, Aib, Nle, inip, Abu, βNal, αNal, Orn, carboxylic acid, andtheir respective D isomers; B¹ is selected from the group consisting ofTrp, βNal, αNal, Leu, Lys, cyclohexylAla, and their respective Disomers; B² is any natural L-amino acid, Pal (3-pyridyl alanine),cycloAla, Aib, Nle, inip, Abu, βNal, αNal, Orn, and their respective Disomers; and isomers, derivatives, prodrugs, metabolite, orpharmaceutically acceptable salts thereof.

The compounds of formulae (I)-(VI) are described in the U.S. ProvisionalPatent App. No. 60/795,960, filed Apr. 28, 2007 and Int. Pat. App. Pub.No.: PCT/US2007/010389, contents of both of which are hereinincorporated by reference.

In some embodiments, the ghrelin receptor antagonist is a peptidecomprising the amino acid sequence of formula (VII):A¹¹-A¹²-A¹³-Gly-Ser-A¹⁴-Phe-Leu-A¹⁵-A¹⁶-A¹⁷-A¹⁸ (SEQ ID NO: 93), whereineach of A¹¹, A¹², and A¹³, is absent, an amino acid, or an aminoprotecting group; A¹⁵, A¹⁶, A¹⁷, and A¹⁸ is independently absent or anamino acid; A¹⁴ is a serine conjugated with a —C(O)C₁-C₂₀ alky group onthe side chain OH of said serine or a diaminopropionic acid conjugatedwith a —C(O)C₁-C₂₀ alky group on one of the amino group diaminopropionicacid, provided at least one of A¹¹, A¹², or A¹³ is present.

In some embodiments, each of A¹¹, A¹², A¹³, A¹⁵, A¹⁶, A¹⁷, and A¹⁸ isindependently selected from the group consisting of alanine, arginine,asparagine, aspartic acid, cysteine, glutamic acid, glutamine, glycine,histidine, isoleucine, leucine, lysine, methionine, phenylalanine,proline, serine, threonine, tryptophan, tyrosine, valine, homocysteine,phosphoserine, phosphothreonine, phosphotyrosine, hydroxyproline,gamma-carboxyglutamate; hippuric acid, octahydroindole-2-carboxylicacid, statine, 1,2,3,4,-tetrahydroisoquinoline-3-carboxylic acid,penicillamine (3-mercapto-D-valine), ornithine, citruline,alpha-methyl-alanine, para-benzoylphenylalanine,para-aminophenylalanine, p-fluorophenylalanine, phenylglycine,propargylglycine, sarcosine, and tert-butylglycine), diaminobutyricacid, 7-hydroxy-tetrahydroisoquinoline carboxylic acid, naphthylalanine,biphenylalanine, cyclohexylalanine, amino-isobutyric acid, norvaline,norleucine, tert-leucine, tetrahydroisoquinoline carboxylic acid,pipecolic acid, phenylglycine, homophenylalanine, cyclohexylglycine,dehydroleucine, 2,2-diethylglycine, 1-amino-1-cyclopentanecarboxylicacid, 1-amino-1-cyclohexanecarboxylic acid, amino-benzoic acid,amino-naphthoic acid, gamma-aminobutyric acid, difluorophenylalanine,nipecotic acid, alpha-amino butyric acid, thienyl-alanine,t-butylglycine, desamino-Tyr, aminovaleric acid, pyroglutaminic acid,alpha-aminoisobutyric acid, gamma-aminobutyric acid, alpha-aminobutyricacid, alpha,gamma-aminobutyric acid, pyridylalanine, α-naphthylalanine,β-naphthylalanine, Ac-β-naphthylalanine, N^(ε)-picoloyl-lysine,4-halo-Phenyl, 4-pyrolidylalanine, isonipecotic acid, and isomers,analogs and derivatives thereof. One of skill in the art would know thatthis definition includes, D- and L-amino acids, alpha- and beta-aminoacids, chemically modified amino acids, naturally occurringnon-proteogenic amino acids, rare amino acids, and chemicallysynthesized compounds that have properties known in the art to becharacteristic of an amino acid. Furthermore, as used herein, the term“amino acid” includes compounds which depart from the structure of thenaturally occurring amino acids, but which have substantially thestructure of an amino acid, such that they can be substituted within apeptide which retains is activity, e.g., biological activity. Thus, forexample, in some embodiments amino acids can also include amino acidshaving side chain modifications or substitutions, and also includerelated organic acids, amides or the like. Without limitation, an aminoacid can be a proteogenic or non-proteogenic amino acid. As used herein,the term “proteogenic” indicates that the amino acid can be incorporatedinto a protein in a cell through well-known metabolic pathways.

The term “alkyl” refers to saturated non-aromatic hydrocarbon chainsthat may be a straight chain or branched chain, containing the indicatednumber of carbon atoms (these include without limitation propyl, allyl,or propargyl), which may be optionally inserted with N, O, or S. Forexample, C₁-C₆ indicates that the group may have from 1 to 6 (inclusive)carbon atoms in it. As used herein, the alkyl can also comprise 1, 2, or3 double and/or triple bonds.

Exemplary amino-protecting groups include, but are not limited to,carbamate protecting groups, such as 2-trimethylsilylethoxycarbonyl(Teoc), 1-methyl-1-(4-biphenylyl)ethoxycarbonyl (Bpoc), t-butoxycarbonyl(BOC), allyloxycarbonyl (Alloc), 9-fluorenylmethyloxycarbonyl (Fmoc),and benzyloxycarbonyl (Cbz); amide protecting groups, such as formyl,acetyl, trihaloacetyl, benzoyl, and nitrophenylacetyl; sulfonamideprotecting groups, such as 2-nitrobenzenesulfonyl; and imine and cyclicimide protecting groups, such as phthalimido and dithiasuccinoyl.Further amino protecting groups, as well as other representativeprotecting groups, are disclosed in Greene and Wuts, Protective Groupsin Organic Synthesis, Chapter 2, 2d ed., John Wiley & Sons, New York,1991, and Oligonucleotides And Analogues A Practical Approach, Ekstein,F. Ed., IRL Press, N.Y, 1991.

In some embodiments of compounds of formula (VII), the alkyl is aC₁-C₁₆alkyl.

In some embodiments of compounds of formula (VII), the alkyl is methyl,ethyl, butyl, pentyl, hexyl, heptyl, octyl, nonyl, decyl, undecyl, ordodecyl.

Without wishing to be bound by theory, addition of at least one of A¹¹,A¹², or A¹³ on the N-terminus of the peptides of formula (VII) block theN-terminal glycine of the ghrelin pentapetide which is necessary forghrelin activity.

In some embodiments, A⁵ is octanoylated serine. In some embodiments, A⁵is an octyanolyatd diaminopropionic acid. In some embodiments, A⁵ is notpalmitolyted diaminopropionic acid, i.e., a diaminopropionic acidconjugated with —C(O)C₁₇alkyl on one of the amino groups of thediaminopropionic acid.

In some embodiments, the peptide of formula (VII) comprises at least one(e.g., 1, 2, 3, 4, 5, 6, 7, 8, 9, 10 or more) D-amino acid. In someembodiments, at least one (e.g., 1, 2, 3, 4, 5, 6, or 7) of A¹¹, A¹²,A¹³, A¹⁵, A¹⁶, A¹⁷, and A¹⁸ is a D-amino acid or derivative thereof. TheD-amino acid can be present at any position in the peptide of formula(VII), for example reading from the N-terminal at position 1, 2, 3, 4,5, 6, 7, 8, 9, 10, 11, or 12. When more than one D-amino acids arepresent, they can be positioned next to or not next to each other.

In some embodiments, the peptide of formula (VII) comprises at least onebeta-amino acid. In some embodiments, at least one (e.g., 1, 2, 3, 4, 5,6, or 7) of A¹¹, A¹², A¹³, A¹⁵, A¹⁶, A¹⁷, A¹⁸ is a beta-amino acid. Thebeta-amino acid can be present at any position in the peptide of formula(VII), for example reading from the N-terminal at position 1, 2, 3, 4,5, 6, 7, 8, 9, 10, 11, or 12. When more than one beta-amino acids arepresent, they can be positioned next to or not next to each other.

In some embodiments, the peptide of formula (VII) comprises at least one(e.g., 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, or 11) peptide bond replaced by alinkage selected from the group consisting of reduced psi peptide bond,urea, thiourea, carbamate, sulfonyl urea, trifluoroethylamine,ortho-(aminoalkyl)-phenylacetic acid, para-(aminoalkyl)-phenylaceticacid, meta-(aminoalkyl)-phenylacetic acid, thioamide, tetrazole, boronicester, and olefinic group. The peptide replacement linkage can bepresent at any position in the peptide of formula (VII), for examplereading from the N-terminal at position 1, 2, 3, 4, 5, 6, 7, 8, 9, 10,or 11. When more than peptide replacement linkages are present, they canbe positioned next to (e.g., on both sides of a given amino acid) or notnext to each other (e.g., only one side of a given amino acid is linkedvia a peptide replacement linkage to the next amino acid).

In some embodiments, the peptide of formula (VII) isGly-Met-Ala-Gly-Ser-(Dap-Oct)-Phe-Leu-Ser-Pro-Glu-His-NH₂ (SEQ ID NO:3).

In some embodiments, the peptide of formula (VII) isGly-Met-Ala-Gly-Ser-(Dap-Palmityl)-Phe-Leu-Ser-Pro-Glu-His-NH₂ (SEQ IDNO: 4).

In some embodiments, the peptide of formula (VII) is notGly-Met-Ala-Gly-Ser-(Dap-Palmityl)-Phe-Leu-Ser-Pro-Glu-His-NH₂ (SEQ IDNO: 4).

Without wishing to be bound by theory, the ghrelin receptor antagonistcan be a dipeptide, a tripeptide, and/or a tetrapeptide. Examples ofsuch peptides include, but are not limited to, the followingembodiments.

In some embodiments, the ghrelin receptor antagonist comprises atetrapeptide core of DTrp-Ala-Trp-DPhe. In some embodiments of this, theposition 1 of the tetrapeptide core comprise an amino acid selected fromthe group consisting of D-Trp, D-β-Nal, D-α-Nal, D-Phe, andD-cyclohexylAla, while the position 2 of the tetrapeptide core compriseL- or D-natural, unnatural, or derivatized amino acid residuesincluding, but not limited to, D-Lys, D-Orn, Ser, D-Pal, D-Leu, and Phe.In some embodiments, the tetrapeptide core comprises 1, 2, 3, 4, or 5additional amino acids at the N-terminus, C-terminus, or both the N- andC-termini of the tetrapeptide core. Without limitation, the N-terminusadditions may comprise any of L- or D-amino acids, such as, but notlimited to, Tyr, His, desamino Tyr, Glyψ, Alaψ, D-Ala, β-Ala, α-Abu,γ-Abu, αγ-Abu, D-Lys, <Glu, D-Arg, D-Orn, carboxylic acid, or mono, ditripeptides or longer peptides such as His-Lys, His-DLys, and DHis-Lys.Without limitation, the C-terminus additions may comprise of any L- orD-natural or unnatural amino acid residues with terminal amidation orcarboxylation including mono, di, tripeptides or longer peptides such asLys, Arg, Lys-Gln, and Lys-Gln-Gly. Examples of tetrapeptide corecontaining antagonist are DHis-DTrp-DPro-DIleNH₂ (SEQ ID NO: 90),DHis-DTrp-DPro-DArgNH₂ (SEQ ID NO: 91), and DβNal-DTrp-DPro-DArgNH₂ (SEQID NO: 92).

In some embodiments, the ghrelin receptor antagonist comprises atripeptide core of DAla-DTrp-Phe. The positions 1 and 2 of thistripeptide core can be substituted with any amino acids selected fromD-amino acid residues or unnatural amino acid residues. The residue atposition 3 can be substituted with an amino acid selected from the groupconsisting of Trp, Leu, Val, Ile, Pro, Phe, cyclohexylAla andcyclopentylAla. In some embodiments, the tetrapeptide core comprises 1,2, 3, 4, or 5 additional amino acids at the N-terminus, C-terminus, orboth the N- and C-termini of the tripeptide core. Without limitation,the N terminus additions can comprise any L- or D-natural, unnaturalamino acid residue, organic carboxylic acids, or dipeptides or longerpeptides with L- or D-natural or unnatural amino acid residues invarious combinations and/or sequences. Some exemplary N-terminusadditions include, but are not limited to, DAla-DβNal, DAla-DαNal,βAla-Trp, His-Trp, DHis-Trp, DHis-DTrp, His-DTrp. Similarly, the Cterminus additions can comprise, without limitation, any L- or D-naturalor unnatural amino acid residues with terminal amidation orcarboxylation including. Exemplary C-terminus additions include, but arenot limited to, mono, di, tripeptides, or longer peptides such as Lys,Arg, Lys-Gln, Lys-Gln-Gly or carboxylic acid. Examples of tripeptidecore containing antagonist are αAib-DTrp-cyclohexylDAlaNH₂(SEQ ID NO:69), AcDTrp-DTrpLysNH₂ (SEQ ID NO: 56) and AcDβNal-Leu-ProNH₂ (SEQ IDNO: 58).

In some embodiments, the ghrelin receptor antagonist comprises thetripeptide core of DTrp-Phe-DTrp. The positions 1 and 3 of this core maybe substituted with amino acids selected from DβNal, DαNal, DPhe, andDcyclohexylAla, and the position 2 of the tripeptide core may besubstituted with amino acids selected from Trp βNal, αNal, Leu, DLeu,and DLys. In one embodiment, there may be additional amino acids at theN-terminus alone, C-terminus alone, or at both N- and C-terminus of thetripeptide core. N terminus additions may comprise a L or D natural orunnatural amino acid residues or organic carboxylic acid. N terminusadditions may comprise a dipeptides, tripeptide, or tetrapeptidescomprising L or D natural or unnatural amino acid residues in variouscombinations and/or sequences, for example DAla-DβNal, DAla-DαNal,βAla-Trp, His-Trp, DHis-Trp, DHis-DTrp, His-DTrp. C terminus additionsmay comprise an amino acid residue selected from Leu, Lys, and Arg. Cterminus additions may also comprise a dipeptide or tripeptidecomprising the amino acids Leu, Lys, Arg, and combinations thereof.Examples of C terminus additions are Leu, Leu-Leu, Leu-Lys, Leu-Arg,Leu-Leu-Lys.

In yet another embodiment, the ghrelin receptor antagonist has at leasta dipeptide core of DTrp-DTrp wherein the core amino acids may besubstituted with D natural and/or derivatized amino acid residues ofTrp, βNal, αNal, Phe, and various combinations thereof. In oneembodiment, there may be additional amino acids at the N-terminus alone,C-terminus alone, or at both the N- and C-terminus of the dipeptidecore. N terminus additions may comprise L or D natural or derivatizedamino acid residues and/or organic carboxylic acid. Additionally, Nterminus additions may also comprise a dipeptide or tripeptide ortetrapeptide or pentapeptide comprising amino acid residues selectedfrom Tyr, His, desamino Tyr, Lys, Glyψ, Alaψ, DAla, αγAbu, αAbu, γAbu,Lys, DLys, isonipecotic carboxylic acid (inip),βAla, DAla, DLys, DThr,DVal, DLeu, cyclohexylAla, cyclopentylAla, DTrp, iPrLys, and diethylguanidinoArg. Examples of N-terminus additions include His-Trp,DLys-Tyr, βAla-Trp, N-AcDLys-Tyr, βAla-Pal, αγAbu-Trp, γAbu-Trp,Ava-Trp, αAbu-Trp, His-DLys, Lys-DHis, DLys-Tyr-DHis. C terminusadditions may comprise L- or D-natural or derivatized amino acidresidues or organic carboxylic acids. C terminus additions may alsocomprise a dipeptide or tripeptide or tetrapeptide or pentapeptidecomprising amino acids residues selected from Phe, DPro, Leu, Met, Ser,Lys, Orn, Arg, cyclohexylAla, and cyclopentylAla. examples of theC-terminus additions are Phe-Lys, DPro-Lys, Phe-DPro-Lys, DPro-Arg,Phe-Met, Phe-Ala, Phe-Ser. Examples of dipeptide core containingantagonist are AcDβNal-DTrpNH₂ (SEQ ID NO: 68) andDThr-DαNal-DTrp-DPro-ArgNH₂ (SEQ ID NO: 48).

In some embodiments, the ghrelin receptor antagonist and/or growthhormone secretatogue antagonist in any of the methods described hereincomprises a compound having a formula I, II, III, IV, V, or VI describedherein. In other embodiments, the ghrelin receptor antagonist and/orgrowth hormone secretatogue antagonist in any of the methods describedherein comprises analogues, prodrug, metabolite, or pharmaceutical saltsthereof of formula I, II, III, IV, V, or VI.

In one embodiment, the ghrelin receptor antagonist isHisDβNalDLysTrpDPheLysNH₂ (SEQ ID NO: 26), or analogues, prodrug,metabolite, or pharmaceutical salts thereof.

In one embodiment, the ghrelin receptor antagonist isHisDTrpDLysTrpDPheLys NH₂ (SEQ ID NO: 16).

In another embodiment, the compounds of the formula (I), excludesHisDβNalDLysTrpDPheLysNH₂ (SEQ ID NO: 26).

In one embodiment, D-Arg1,D-Phe5,D-Trp7,9,Leu11 substance P functions asa low-potency antagonist but a high-potency full inverse agonist on theghrelin receptor.

In some embodiments, the ghrelin receptor antagonists described herein,either alone or in combination with a GOAT inhibitor, inhibit GHsecretion in a subject. The GH secretion can be inhibited by at least5%, at least 10%, at least 20%, at least 30%, at least 40%, at least50%, at least 60%, at least 70%, at least 80%, at least 90%, at least95% or complete inhibition relative to an untreated control. Skilledartisan is well aware of the methods for measuring GH secretion in vivo,for example the methods described in the Examples section.

In some embodiments, the ghrelin receptor antagonists described herein,either alone or in combination with a GOAT inhibitor, decrease orinhibit food intake in a subject by at least 5%, at least 10%, at least20%, at least 30%, at least 40%, at least 50%, at least 60%, at least70%, at least 80%, at least 90%, at least 95% or complete inhibitionrelative to an untreated control. As used herein, food intake refers tocaloric intake by a subject. In some cases, food intake also refers toweight of food consumed by a subject.

In one embodiment, the ghrelin receptor antagonist and/or growth hormonesecretatogue antagonist in any of the methods described herein comprisescompositions of compounds having formulae I, II, III, IV, V, VI, VII, orcombinations thereof. For example, compositions of compounds havingformulae I, II, III, IV, V, VI, VII, or combinations thereof are usedfor the treatment of obesity or obesity related diseases and disordersin a subject, and can be combined with a surgical or mechanicalprocedure used to treat obesity or other obesity treatment. Suchprocedures include but are not limited to gastric bypass surgery andgastric banding.

In one embodiment, the ghrelin receptor antagonist and/or growth hormonesecretatogue antagonist in any of the methods described herein consistsessentially of compositions of compounds having formulae I, II, III, IV,V, VI, VII, or combinations thereof.

In one embodiment, the ghrelin receptor antagonist and/or growth hormonesecretatogue antagonist in any of the methods described herein consistsof compositions of compounds having formulae I, II, III, IV, V, VI, VII,or combinations thereof.

In one embodiment, compositions of compounds having formulae I, II, III,IV, V, VI, VII, or combinations thereof can be used in combination forthe treatment of various diseases and disorders described herein, e. g.cancer treatment, obesity treatment, metabolic syndrome and diabetestreatment. These treatments are well known to physicians skilled in theart.

The experimental data regarding the effect of the compounds of thisinvention has been produced in well known animal models that aretypically used for the effects of anti-obesity treatments at the firststage and are thus likely to be applicable to human obesity.

In some embodiments, the individual is obese or overweight. Typically,an overweigh individual is considered to have a body mass index (BMI) ofover 25 but under 30 and an obese individual is considered to have a BMIof over 30.

In one embodiment, the methods and uses of the present invention areapplicable for both sexes and all age groups including children andteenagers.

In some embodiments, the GOAT inhibitor and/or ghrelin receptorantagonist described herein are administered to a subject in need ofsuch treatment for any of the disease or disorders or associateddisorders discussed and described herein, e. g. obesity, overeating,diabetes mellitus, metabolic syndrome, hypertension, elevated plasmainsulin concentrations, insulin resistance, dyslipidemias, andhyperlipidemia, unregulated cellular proliferation, and cancer such ashormonally functional endocrine or non-endocrine tumors, breast andprostate cancer, ectopic neuroendocrine tumors, carcinoid and pancreatictumors, and prostate cancer, osteoblast cancer, pancreatic cancer,adenocarcinomas and hepatoma. In one embodiment, a subject who is inneed of treatment is one who has been clinically diagnosed with adisease, disorders or associated disorders described herein by methodswell known to one skilled in the art, e. g. a physician. In anotherembodiment, a subject who is in need of treatment is one who is at riskof developing the disease, disorders or associated disorders describedherein. For example, a subject who has had a sudden gain of weight in ashort period of time, e. g. gaining 5 kilograms, 10 kilograms, 20kilograms, 30 kilograms, 40 kilograms, or 50 kilogram within a period of1 month, 2 months, 3 months, or four months. Without wishing to be boundby theorym, such a subject is at high risk for developing diabetesmellitus, metabolic syndrome, hypertension, elevated plasma insulinconcentrations, insulin resistance, dyslipidemias, and hyperlipidemiaand other obesity related disorders.

As used herein, a “subject” means a human or animal. Usually the animalis a vertebrate such as a primate, rodent, domestic animal or gameanimal. Primates include chimpanzees, cynomologous monkeys, spidermonkeys, and macaques, e.g., Rhesus. Rodents include mice, rats,woodchucks, ferrets, rabbits and hamsters. Domestic and game animalsinclude cows, horses, pigs, deer, bison, buffalo, feline species, e.g.,domestic cat, canine species, e.g., dog, fox, wolf, avian species, e.g.,chicken, emu, ostrich, and fish, e.g., trout, catfish and salmon.Patient or subject includes any subset of the foregoing, e.g., all ofthe above, but excluding one or more groups or species such as humans,primates or rodents. In certain embodiments, the subject is a mammal,e.g., a primate, e.g., a human. The terms, “patient” and “subject” areused interchangeably herein.

In some embodiments of the methods described herein, the method furthercomprising selecting a subject identified as being in need of treatmentby a GOAT inhibitor and a ghrelin antagonit. A subject suffering from adisease or disorder can be selected based on the symptoms presented asdescribed herein.

In some embodiments, the subject is a mammal. In other embodiments, thesubject is an animal, e. g. a cat, a dog, a horse. In one preferredembodiment, the subject is a human.

DEFINITIONS

Unless stated otherwise, or implicit from context, the following termsand phrases include the meanings provided below. Unless explicitlystated otherwise, or apparent from context, the terms and phrases belowdo not exclude the meaning that the term or phrase has acquired in theart to which it pertains. The definitions are provided to aid indescribing particular embodiments, and are not intended to limit theclaimed invention, because the scope of the invention is limited only bythe claims. Further, unless otherwise required by context, singularterms shall include pluralities and plural terms shall include thesingular.

As used herein the term “comprising” or “comprises” is used in referenceto compositions, methods, and respective component(s) thereof, that areessential to the invention, yet open to the inclusion of unspecifiedelements, whether essential or not.

As used herein the term “consisting essentially of” refers to thoseelements required for a given embodiment. The term permits the presenceof additional elements that do not materially affect the basic and novelor functional characteristic(s) of that embodiment of the invention.

The term “consisting of” refers to compositions, methods, and respectivecomponents thereof as described herein, which are exclusive of anyelement not recited in that description of the embodiment.

The singular terms “a,” “an,” and “the” include plural referents unlesscontext clearly indicates otherwise. Similarly, the word “or” isintended to include “and” unless the context clearly indicatesotherwise.

Although methods and materials similar or equivalent to those describedherein can be used in the practice or testing of this disclosure,suitable methods and materials are described below. The term “comprises”means “includes.” The abbreviation, “e.g.” is derived from the Latinexempli gratia, and is used herein to indicate a non-limiting example.Thus, the abbreviation “e.g.” is synonymous with the term “for example.”

The terms “decrease”, “reduced”, “reduction”, “decrease” or “inhibit”are all used herein generally to mean a decrease by a statisticallysignificant amount. However, for avoidance of doubt, “reduced”,“reduction” or “decrease” or “inhibit” means a decrease by at least 10%as compared to a reference level, for example a decrease by at leastabout 20%, or at least about 30%, or at least about 40%, or at leastabout 50%, or at least about 60%, or at least about 70%, or at leastabout 80%, or at least about 90% or up to and including a 100% decrease(e.g. absent level as compared to a reference sample), or any decreasebetween 10-100% as compared to a reference level.

The terms “increased”, “increase” or “enhance” or “activate” are allused herein to generally mean an increase by a statically significantamount; for the avoidance of any doubt, the terms “increased”,“increase” or “enhance” or “activate” means an increase of at least 10%as compared to a reference level, for example an increase of at leastabout 20%, or at least about 30%, or at least about 40%, or at leastabout 50%, or at least about 60%, or at least about 70%, or at leastabout 80%, or at least about 90% or up to and including a 100% increaseor any increase between 10-100% as compared to a reference level, or atleast about a 2-fold, or at least about a 3-fold, or at least about a4-fold, or at least about a 5-fold or at least about a 10-fold increase,or any increase between 2-fold and 10-fold or greater as compared to areference level.

The term “statistically significant” or “significantly” refers tostatistical significance and generally means a two standard deviation(2SD) below normal, or lower, concentration of the marker. The termrefers to statistical evidence that there is a difference. It is definedas the probability of making a decision to reject the null hypothesiswhen the null hypothesis is actually true. The decision is often made us

Unless otherwise noted, the abbreviations for the residues of aminoacids used herein are in agreement with the standard nomenclature; andare as follows: Gly: Glycine; Tyr: Tyrosine; Ile: Isoleucine; Glu:Glutamic Acid; Thr: Threonine; Phe: Phenylalanine; Ala: Alanine; Lys:Lysine; Asp: Aspartic Acid; Cys: Cysteine; Arg: Arginine; Gln:Glutamine; Pro: Proline; Leu: Leucine; Met: Methionine; Ser: Serine;Asn: Asparagine; His: Histidine; Trp: Tryptophan; Val: Valine; Orn:Ornithine; Desamino-Tyr: Desamino Tyrosine; Desamino-His: DesaminoHistidine; Desamino-alpha-Aib: Desamino-alpha-aminoisobutyric acid;Desamino-alpha-Abu: Desamin-alpha-aminobutyric acid;Desamino-alpha-gamma-Abu (Desamino-α,γ-Abu):Desamino-alpha-gamma-aminobutyric acid.

Moreover, all of the three letter-abbreviations of the amino acidspreceded by a “D” indicates the dextro-isomer of the amino acid residue.Glycine is considered to be included in the term naturally occurringL-amino acids. Other abbreviations used herein include: Aib:aminoisobutyric acid; inip: isonipecotyl; Abu-aminobutyric acid;alpha-Nal: alpha-naphthylalanine; beta-Na: beta-naphthylalanine; D-alphaNal: alpha-naphthyl-D-alanine; D-beta-Nal: beta-naphthyl-D-alanine; Pal:3-pyridyl alanine; CHx: cyclohexyl; CHxAla: L-cyclohexylalanine; Ava:Aminovaleric acid; IMA: N-alpha-imidazole acetic acid; imc: imidazolecarboxylic acid; beta-Ala: beta-Alanine; ILys: Lysine (iPr) which isisopropyl-α-N^(ε)lysine; α,γ-Abu: alpha-gamma-diaminobutyric acid; Nle:norleucine; PicLys: N^(ε)-picoloyl-lysine); inip: isonipecotoccarboxylic acid; NMePhe: N-methylated phenylalanine; Sar: sarcosine(N-methylglycine); <Glu: pyroglutamic acid; Ac-D-β-Nal: acetylatedD-beta-naphthylalanine; Ac-D-α-Nal: acetylated D-alpha-naphthylalanine;and N-Ac-D-beta-Nal: N-acetyl D-β-Naphthylalanine.

The terms “administration of” and or “administering” a compound shouldbe understood to mean providing a ghrelin receptor antagonist compoundof the invention, a prodrug or an active metabolite of a compound of theinvention to a subject in need of treatment. As such, the term“administer” refers to the placement of a composition into a subject bya method or route which results in at least partial localization of thecomposition at a desired site such that GOAT is inhibited and/or ghrelinreceptor is antagonized. Compounds described herein can be administeredby any appropriate route known in the art including, but not limited tooral or parenteral routes, including intravenous, intramuscular,subcutaneous, transdermal, airway (aerosol), pulmonary, nasal, rectal,and topical (including buccal and sublingual) administration.

Exemplary modes of administration include, but are not limited to,injection, infusion, instillation, inhalation, or ingestion. “Injection”includes, without limitation, intravenous, intramuscular, intraarterial,intrathecal, intraventricular, intracapsular, intraorbital,intracardiac, intradermal, intraperitoneal, transtracheal, subcutaneous,subcuticular, intraarticular, sub capsular, subarachnoid, intraspinal,intracerebro spinal, and intrasternal injection and infusion. Inpreferred embodiments, the compositions are administered by intravenousinfusion or injection.

The term “ghrelin receptor” as used herein includes growth hormonesecretagogue receptor, GHS-R1a and subtypes, isoforms and variantsthereof.

The term “diabetes,” as used herein, includes both insulin-dependentdiabetes mellitus (i.e., IDDM, also known as type I diabetes) andnon-insulin-dependent diabetes mellitus (i.e., NIDDM, also known as TypeII diabetes). Type I diabetes, or insulin-dependent diabetes, is theresult of an absolute deficiency of insulin, the hormone which regulatesglucose utilization. Type II diabetes, or insulin-independent diabetes(i.e., non-insulin-dependent diabetes mellitus), often occurs in theface of normal, or even elevated levels of insulin and appears to be theresult of the inability of tissues to respond appropriately to insulin.Most of the Type II diabetics are also obese. The compositions of thepresent invention are useful for treating both Type I and Type IIdiabetes. The compositions are especially effective for treating Type IIdiabetes. The compounds or combinations of the present invention arealso useful for treating and/or preventing gestational diabetesmellitus.

There are two forms of Diabetes mellitus: (1) insulin dependent or Type1 Diabetes (a.k.a., Juvenile Diabetes, Brittle Diabetes, InsulinDependent Diabetes Mellitus (IDDM)) and (2) non-insulin-dependent orType II Diabetes (a.k.a., NIDDM). Type 1 Diabetes develops most often inyoung people but can appear in adults. Type 2 Diabetes develops mostoften in middle aged and older adults, but can appear in young people.Diabetes is a disease derived from multiple causative factors andcharacterized by elevated levels of plasma glucose (hyperglycemia) inthe fasting state or after administration of glucose during an oralglucose tolerance test. A decrease in β-cell mass occurs in both Type 1and Type 2 Diabetes.

The terms “diabetes” and “diabetes mellitus” are used interchangeablyherein. The World Health Organization defines the diagnostic value offasting plasma glucose concentration to 7.0 mmol/1 (126 mg/dl) and abovefor Diabetes Mellitus (whole blood 6.1 mmol/1 or 110 mg/dl), or 2-hourglucose level≧11.1 mmol/L (≧200 mg/dL). Other values suggestive of orindicating high risk for Diabetes Mellitus include elevated arterialpressure≧140/90 mm Hg; elevated plasma triglycerides (≧1.7 mmol/L; 150mg/dL) and/or low HDL-cholesterol (<0.9 mmol/L, 35 mg/dl for men; <1.0mmol/L, 39 mg/dL women); central obesity (males: waist to hipratio >0.90; females: waist to hip ratio >0.85) and/or body mass indexexceeding 30 kg/m²; microalbuminuria, where the urinary albuminexcretion rate ≧20 μg/min or albumin:creatinine ratio ≧30 mg/g).

A “pre-diabetic condition” refers to a metabolic state that isintermediate between normal glucose homeostasis, metabolism, and statesseen in Diabetes Mellitus. Pre-diabetic conditions include, withoutlimitation, Metabolic Syndrome (“Syndrome X”), Impaired GlucoseTolerance (IGT), and Impaired Fasting Glycemia (IFG). IGT refers topost-prandial abnormalities of glucose regulation, while IFG refers toabnormalities that are measured in a fasting state. The World HealthOrganization defines values for IFG as a fasting plasma glucoseconcentration of 6.1 mmol/L (100 mg/dL) or greater (whole blood 5.6mmol/L; 100 mg/dL), but less than 7.0 mmol/L (126 mg/dL)(whole blood 6.1mmol/L; 110 mg/dL). Metabolic Syndrome according to National CholesterolEducation Program (NCEP) criteria are defined as having at least threeof the following: blood pressure 130/85 mm Hg; fasting plasma glucose≧6.1 mmol/L; waist circumference >102 cm (men) or >88 cm (women);triglycerides ≧1.7 mmol/L; and HDL cholesterol <1.0 mmol/L (men) or 1.3mmol/L (women).

“Impaired glucose tolerance” (IGT) is defined as having a blood glucoselevel that is higher than normal, but not high enough to be classifiedas Diabetes Mellitus. A subject with IGT will have two-hour glucoselevels of 140 to 199 mg/dL (7.8 to 11.0 mmol) on the 75 g oral glucosetolerance test. These glucose levels are above normal but below thelevel that is diagnostic for Diabetes. Subjects with impaired glucosetolerance or impaired fasting glucose have a significant risk ofdeveloping Diabetes and thus are an important target group for primaryprevention.

“Normal glucose levels” is used interchangeably with the term“normoglycemic” and refers to a fasting venous plasma glucoseconcentration of less than 6.1 mmol/L (110 mg/dL). Although this amountis arbitrary, such values have been observed in subjects with provennormal glucose tolerance, although some may have IGT as measured by oralglucose tolerance test (OGTT). A baseline value, index value, orreference value in the context of the present invention and definedherein can comprise, for example, “normal glucose levels.”

In general, treatment of Diabetes is determined by standard medicalmethods. A goal of Diabetes treatment is to bring sugar levels down toas close to normal as is safely possible. Commonly set goals are 80-120milligrams per deciliter (mg/dl) before meals and 100-140 mg/dl atbedtime. A particular physician may set different targets for thepatent, depending on other factors, such as how often the patient haslow blood sugar reactions. Useful medical tests include tests on thepatient's blood and urine to determine blood sugar level, tests forglycosylated hemoglobin level (HbAlc; a measure of average blood glucoselevels over the past 2-3 months, normal range being 4-6%), tests forcholesterol and fat levels, and tests for urine protein level. Suchtests are standard tests known to those of skill in the art (see, forexample, American Diabetes Association, 1998). A successful treatmentprogram can also be determined by having fewer patients in the programwith complications relating to Diabetes, such as diseases of the eye,kidney disease, or nerve disease.

Type 1 Diabetes is an autoimmune disease that results in destruction ofinsulin-producing beta cells of the pancreas. Lack of insulin causes anincrease of fasting blood glucose (around 70-120 mg/dL in nondiabeticpeople) that begins to appear in the urine above the renal threshold(about 190-200 mg/dl in most people). Type 1 Diabetes can be diagnosedusing a variety of diagnostic tests that include, but are not limitedto, the following: (1) glycated hemoglobin (AlC) test, (2) random bloodglucose test and/or (3) fasting blood glucose test.

The Glycated hemoglobin (AlC) test is a blood test that reflects theaverage blood glucose level of a subject over the preceding two to threemonths. The test measures the percentage of blood glucose attached tohemoglobin, which correlates with blood glucose levels (e.g., the higherthe blood glucose levels, the more hemoglobin is glycosylated). An AlClevel of 6.5 percent or higher on two separate tests is indicative ofDiabetes. A result between 6 and 6.5 percent is considered prediabetic,which indicates a high risk of developing Diabetes.

The Random Blood Glucose Test comprises obtaining a blood sample at arandom time point from a subject suspected of having Diabetes. Bloodglucose values can be expressed in milligrams per deciliter (mg/dL) ormillimoles per liter (mmol/L). A random blood glucose level of 200 mg/dL(11.1 mmol/L) or higher indicates the subject likely has Diabetes,especially when coupled with any of the signs and symptoms of Diabetes,such as frequent urination and extreme thirst.

For the fasting blood glucose test, a blood sample is obtained after anovernight fast. A fasting blood glucose level less than 100 mg/dL (5.6mmol/L) is considered normal. A fasting blood glucose level from 100 to125 mg/dL (5.6 to 6.9 mmol/L) is considered prediabetic, while a levelof 126 mg/dL (7 mmol/L) or higher on two separate tests is indicative ofDiabetes.

Type 1 Diabetes can also be distinguished from type 2 Diabetes using aC-peptide assay, which is a measure of endogenous insulin production.The presence of anti-islet antibodies (to Glutamic Acid Decarboxylase,Insulinoma Associated Peptide-2 or insulin), or lack of insulinresistance, determined by a glucose tolerance test, is also indicativeof type 1, as many type 2 diabetics continue to produce insulininternally, and all have some degree of insulin resistance.

Testing for GAD 65 antibodies has been proposed as an improved test fordifferentiating between type 1 and type 2 Diabetes as it appears thatthe immune system is involved in Type 1 Diabetes etiology.

The non-obese diabetic (NOD) mouse provides an animal model for thespontaneous development of Type 1 Diabetes. NOD mice develop insulitisas a result of leukocyte infiltration into the pancreatic islet, whichin turn leads to the destruction of pancreatic islets and a Type 1diabetic phenotype (Makino S, et al., (1980) Jikken Dobutsu 29 (1):1-13; Kikutani H, and Makino S (1992) Adv. Immunol. 51: 285-322).

The methods described herein are also useful for treating Type 1Diabetes in a subject.

In the context of type 1 Diabetes, “treating” or “treatment” refers topartial or total inhibition, delay or prevention of the progression oftype 1 Diabetes, pre-diabetic conditions, and complications associatedwith type 1 Diabetes or pre-diabetic conditions; inhibition, delay orprevention of the recurrence of type 2 Diabetes, pre-diabeticconditions, or complications associated with type 1 Diabetes orpre-diabetic conditions; or the prevention of the onset or developmentof type 1 Diabetes, pre-diabetic conditions, or complications associatedwith type 1 Diabetes or pre-diabetic conditions (chemoprevention) in asubject.

In the context of Type 1 Diabetes, “therapeutically effective amount”refers to an amount of GOAT inhibitor and/or ghrelin antagonistadministered to a subject that is sufficient to produce a statisticallysignificant, measurable change in at least one symptom of Type 1Diabetes, such as glycosylated hemoglobin level, fasting blood glucoselevel, hypoinsulinemia, etc. . . . . Efficacy of treatment with apeptide can be assessed by measuring changes in blood glucose and/orinsulin levels or as described below.

The efficacy of a given treatment for Type 1 Diabetes can be determinedby the skilled clinician. However, a treatment is considered “effectivetreatment,” as the term is used herein, if any one or all of the signsor symptoms of Type 1 Diabetes, for example, hyperglycemia are alteredin a beneficial manner, other clinically accepted symptoms or markers ofdisease are improved, or even ameliorated, e.g., by at least 10%following treatment with a peptide as described herein. Efficacy canalso be measured by a failure of an individual to worsen as assessed byhospitalization or need for medical interventions (i.e., progression ofthe disease is halted or at least slowed). Methods of measuring theseindicators are known to those of skill in the art and/or describedherein. Treatment includes any treatment of a disease in an individualor an animal (some non-limiting examples include a human, or a mammal)and includes: (1) inhibiting the disease, e.g., arresting, or slowingthe loss of beta cells; or (2) relieving the disease, e.g., causingregression of symptoms, increasing pancreatic beta cell mass; and (3)preventing or reducing the likelihood of the development of acomplication of Type 1 Diabetes, e.g., diabetic retinopathy.

As used herein, the term “delaying the onset of Type 1 Diabetes” in asubject refers to a delay of onset of at least one symptom of Type 1Diabetes (e.g., hyperglycemia and/or hypoinsulinemia) of at least oneweek, at least 2 weeks, at least 1 month, at least 2 months, at least 6months, at least 1 year, at least 2 years, at least 5 years, at least 10years, at least 20 years, at least 30 years, at least 40 years or more,and can include the entire lifespan of the subject.

Type 2 Diabetes results from a combination of insulin resistance andimpaired insulin secretion but ultimately many people with Type 2Diabetes show markedly reduced pancreatic β-cell mass and functionwhich, in turn, causes Type 2 diabetic persons to have a “relative”deficiency of insulin because pancreatic β-cells are producing someinsulin, but the insulin is either too little or isn't working properlyto adequately allow glucose into cells to produce energy. Recent autopsystudies have shown clear evidence of ongoing β-cell death (apoptosis) inpeople with Type 2 Diabetes. Therefore, therapeutic approaches toprovide more β-cells could provide a significant treatment for reversingor curing Type 2 Diabetes.

Uncontrolled Type 2 Diabetes leads to excess glucose in the blood,resulting in hyperglycemia, or high blood sugar. A person with Type 2Diabetes experiences fatigue, increased thirst, frequent urination, dry,itchy skin, blurred vision, slow healing cuts or sores, more infectionsthan usual, numbness and tingling in feet. Without treatment, a personwith Type 2 Diabetes will become dehydrated and develop a dangerouslylow blood volume. If Type 2 Diabetes remains uncontrolled for a longperiod of time, more serious symptoms may result, including severehyperglycemia (blood sugar over 600 mg) lethargy, confusion, shock, andultimately “hyperosmolar hyperglycemic non-ketotic coma.” Persistent oruncontrolled hyperglycemia is associated with increased and prematuremorbidity and mortality. As such, therapeutic control of glucosehomeostasis, lipid metabolism, obesity, and hypertension are criticallyimportant in the clinical management and treatment of Diabetes mellitus.

The methods of the invention are useful for treating type 2 DiabetesMellitus or a pre-diabetic condition in a subject or preventing type 2Diabetes or pre-diabetic conditions in a subject. Skilled artisan iswell aware that type 2 Diabetes Mellitus is also known as non-insulindependent Diabetes mellitus.

“Complications related to type 2 Diabetes” or “complications related toa pre-diabetic condition” can include, without limitation, diabeticretinopathy, diabetic nephropathy, blindness, memory loss, renalfailure, cardiovascular disease (including coronary artery disease,peripheral artery disease, cerebrovascular disease, atherosclerosis, andhypertension), neuropathy, autonomic dysfunction, hyperglycemichyperosmolar coma, or combinations thereof.

In the context of type 2 Diabetes, “treating” or “treatment” refers topartial or total inhibition, delay or prevention of the progression oftype 2 Diabetes, pre-diabetic conditions, and complications associatedwith type 2 Diabetes or pre-diabetic conditions; inhibition, delay orprevention of the recurrence of type 2 Diabetes, pre-diabeticconditions, or complications associated with type 2 Diabetes orpre-diabetic conditions; or the prevention of the onset or developmentof type 2 Diabetes, pre-diabetic conditions, or complications associatedwith type 2 Diabetes or pre-diabetic conditions (chemoprevention) in asubject.

“Obesity” is a condition in which there is an excess of body fat. Theoperational definition of obesity is based on the Body Mass Index (BMI),which is calculated as body weight per height in meters squared (kg/m²).“Obesity” refers to a condition whereby an otherwise healthy subject hasa Body Mass Index (BMI) greater than or equal to 30 kg/m², or acondition whereby a subject with at least one co-morbidity has a BMIgreater than or equal to 27 kg/m². An “obese subject” is an otherwisehealthy subject with a Body Mass Index (BMI) greater than or equal to 30kg/m² or a subject with at least one co-morbidity with a BMI greaterthan or equal to 27 kg/m². A “subject at risk of obesity” is anotherwise healthy subject with a BMI of 25 kg/m² to less than 30 kg/m²or a subject with at least one co-morbidity with a BMI of 25 kg/m² toless than 27 kg/m².

The term “drug” or “compound” as used herein refers to a chemical entityor biological product, or combination of chemical entities or biologicalproducts, administered to a person to treat or prevent or control adisease or condition.

As used herein, the terms “effective” and “effectiveness” includes bothpharmacological effectiveness and physiological safety. Pharmacologicaleffectiveness refers to the ability of the treatment to result in adesired biological effect in the patient. Physiological safety refers tothe level of toxicity, or other adverse physiological effects at thecellular, organ and/or organism level (often referred to asside-effects) resulting from administration of the treatment. “Lesseffective” means that the treatment results in a therapeuticallysignificant lower level of pharmacological effectiveness and/or atherapeutically greater level of adverse physiological effects. Theghrelin receptor antagonists described herein are effective in treatingobesity and obesity related diseases and disorders including diabetesand various types of cancer.

A “cancer” in an animal refers to the presence of cells possessingcharacteristics typical of cancer-causing cells, such as uncontrolledproliferation, immortality, metastatic potential, rapid growth andproliferation rate, and certain characteristic morphological features.Often, cancer cells will be in the form of a tumor, but such cells mayexist alone within an animal, or may be a non-tumorigenic cancer cell,such as a leukemia cell. In some circumstances, cancer cells will be inthe form of a tumor; such cells may exist locally within an animal, orcirculate in the blood stream as independent cells, for example,leukemic cells. Examples of cancer include but are not limited to breastcancer, a melanoma, adrenal gland cancer, biliary tract cancer, bladdercancer, brain or central nervous system cancer, bronchus cancer,blastoma, carcinoma, a chondrosarcoma, cancer of the oral cavity orpharynx, cervical cancer, colon cancer, colorectal cancer, esophagealcancer, gastrointestinal cancer, glioblastoma, hepatic carcinoma,hepatoma, kidney cancer, leukemia, liver cancer, lung cancer, lymphoma,non-small cell lung cancer, osteosarcoma, ovarian cancer, pancreascancer, peripheral nervous system cancer, prostate cancer, sarcoma,salivary gland cancer, small bowel or appendix cancer, small-cell lungcancer, squamous cell cancer, stomach cancer, testis cancer, thyroidcancer, urinary bladder cancer, uterine or endometrial cancer, andvulval cancer.

The term “inverse agonist” as used herein refers to an agent which bindsto the same ghrelin receptor binding-site as an agonist for thatreceptor but exerts the opposite pharmacological effect, decreaseintracellular IP3 levels.

The term “antagonist” is used in the broadest sense, and includes anymolecule that partially or fully blocks, inhibits, or neutralizes abiological activity of ghrelin.

As used herein, the term “peptide’ refers to a short polymer formed fromthe linking of amino acids. A “peptide” is at least 4, or at least 5amino acids and no more than to 50 amino acids in length.

As used herein, the term “metabolic syndrome” refer to a group ofmetabolic risk factors in one person. They include: abdominal obesity(excessive fat tissue in and around the abdomen), atherogenicdyslipidemia (blood fat disorders—high triglycerides, low HDLcholesterol and high LDL cholesterol—that foster plaque buildups inartery walls), elevated blood pressure, insulin resistance or glucoseintolerance (the body can't properly use insulin or blood sugar),rrothrombotic state (e. g., high fibrinogen or plasminogen activatorinhibitor-1 in the blood), and proinflammatory state (e.g., elevatedC-reactive protein in the blood). There are currently no well-acceptedcriteria for diagnosing the metabolic syndrome. The criteria proposed bythe National Cholesterol Education Program (NCEP) Adult Treatment PanelIII (ATP III), with minor modifications, are currently recommended andwidely used.

Ghrelin O-Acyltransferase (GOAT) Inhibitor

In some embodiments, the methods described herein comprisesadministering an effective amount of ghrelin O-acyltransferase (GOAT)inhibitor.

In one embodiment, the GOAT inhibitor comprises an octanoylated ghrelinpentapeptide, and wherein the octanoylation is at position three of thepentapeptide.

In one embodiment, the octanoylated ghrelin pentapeptide isGly-Ser-[Ser-Octanoyl³]-Phe-Leu (SEQ ID NO: 1). In another embodiment,the octanoylated ghrelin pentapeptide is Gly-Ser-[Dap-Octanoyl³]-Phe-Leu(SEQ ID NO: 2).

Ghrelin is a 28 amino acid, appetite-stimulating peptide hormonesecreted by the food-deprived stomach. Serine-3 of ghrelin is acylatedwith an eight-carbon fatty acid, octanoate, which is required for itsendocrine actions. Ghrelin that is deleted of the octanoate, known asdesoctanoylated ghrelin is biologically inactive. Recently, the membranebound acyltransferase that catalyses the addition of octanoate toSerine-3 was identified (28). The enzyme is named ghrelinO-acyltransferase (GOAT). GOAT was shown to also transfer octanoyl to apentapeptide containing only the N-terminal five amino acids ofproghrelin, the 94 amino acid protein precursor of ghrelin (29). Yang,et, al. 2008 (29) showed that GOAT activity could be inhibited by anoctanoylated ghrelin pentapeptide, and its potency was enhanced 45-foldwhen the octanoylated serine-3 was replaced by octanoylateddiaminopropionic acid.

In some embodiments, the GOAT inhibitors comprises ghrelin pentapeptidesor peptidomimetics that have serine at position three and the serine atposition 3 is not octanoylated. While not wishing to be bound by theory,such desoctanoylated ghrelin pentapeptides or peptidomimetics inhibitsGOAT acylation of ghrelin by serving as decoy substrates to compete withdesoctanoylated ghrelin thereby saturating the GOAT substrate bindingsites.

Other examples of inhibitors that comprise octanoylated ghrelinpentapeptide include, but not limited to, peptides from Phoenixpharmaceutical Inc., e. g. Octanoylated Ghrelin Pentapeptide/Ghrelin(1-5)-Amide, catalog No. 031-41: Gly-Ser-Ser(Octanoyl)-Phe-Leu-NH₂ (SEQID NO: 94); Ghrelin (1-5)-Amide [Ser3(Des-Octanoyl)], catalog No.031-42: Gly-Ser-Ser(Des-Octanoyl)-Phe-Leu-NH₂ (SEQ ID NO: 5); Ghrelin[Dap-Octanyol3], catalog No. 031-58:Gly-Ser-Dap(Octanoyl)-Phe-Leu-Ser-Pro-Glu-His-Gln-Arg-Val-Gln-Gln-Arg-Lys-Glu-Ser-Lys-Lys-Pro-Pro-Ala-Lys-Leu-Gln-Pro-Arg-NH₂(SEQ ID NO: 6); [Dap3] Octanoyl-Ghrelin (1-5) Amide, catalog No.:032-14: Gly-Ser-Dap(Octanoyl)-Phe-Leu-NH₂ (SEQ ID NO: 7); Ghrelin(1-4)-Amide, catalog No. 031-67: Gly-Ser-Ser(Des-Octanoyl)-Phe-NH₂ (SEQID NO: 8); Ghrelin (1-18) [Ser3(Des-Octanoyl)] (Motilin-RelatedPeptide), catalog No.: 031-47:Gly-Ser-Ser(Des-Octanoyl)-Phe-Leu-Ser-Pro-Glu-His-Gln-Arg-Val-Gln-Gln-Arg-Lys-Glu-Ser-NH₂(SEQ ID NO: 9); and Ghrelin [Tyr29] [Ser3(Des-Octanoyl)], catalog No.:031-55:Gly-Ser-Ser(Des-Octanoyl)-Phe-Leu-Ser-Pro-Glu-His-Gln-Arg-Val-Gln-Gln-Arg-Lys-Glu-Ser-Lys-Lys-Pro-Pro-Ala-Lys-Leu-Gln-Pro-Arg-Tyr-NH₂(SEQ ID NO: 10).

Ghrelin Receptor Antagonist and/or Growth Hormone SecretatogueAntagonist Compounds

Ghrelin receptor antagonist and/or growth hormone secretatogueantagonist compounds useful in the methods described herein are fullydescribed in PCT/US2007/10389 which is incorporated herein by referencein its entirety.

In some embodiments, the ghrelin receptor antagonist compounds useful inthe methods described herein includes a compound with the formulaTyr-DTrp-DLys-Trp-DPhe-NH2 (SEQ ID NO: 11), Tyr-DTrp-Lys-Trp-DPhe-NH2(SEQ ID NO: 12), His-DTrp-DLys-Trp-DPhe-NH2 (SEQ ID NO: 13),His-DTrp-DLys-Phe-DTrp-NH2 (SEQ ID NO: 14), His-DTrp-DArg-Trp-DPhe-NH2(SEQ ID NO: 15), His-DTrp-DLys-Trp-DPhe-Lys-NH2 (SEQ ID NO: 16),DesaminoTyr-DTrp-Ala-Trp-DPhe-NH2 (SEQ ID NO: 17),DesaminoTyr-DTrp-DLys-Trp-DPhe-NH2 (SEQ ID NO: 18),DeaminoTyr-DTrp-Ser-Trp-DPhe-Lys-NH2 (SEQ ID NO: 19),DesaminoTyr-DTrp-Ser-Trp-DPhe-NH2 (SEQ ID NO: 20),His-DTrp-DTrp-Phe-Met-NH2 (SEQ ID NO: 21), Tyr-DTrp-DTrp-Phe-Phe-NH2(SEQ ID NO: 22), Glyψ[CH2NH]-DβNal-Ala-Trp-DPhe-Lys-NH2 (SEQ ID NO: 23),Glyψ[CH₂NH]-DβNal-DLys-Trp-DPhe-Lys-NH₂ (SEQ ID NO: 24),DAla-DβNal-DLys-DTrp-Phe-Lys-NH2 (SEQ ID NO: 25),His-DβNal-DLys-Trp-DPhe-Lys-NH2 (SEQ ID NO: 26),Ala-His-DTrp-DLys-Trp-DPhe-Lys-NH2 (SEQ ID NO: 27),Alaψ[CH2NH]-DβNal-Ala-Trp-DPhe-Lys-NH2 (SEQ ID NO: 28),DβNal-Ala-Trp-DPhe-Ala-NH2 (SEQ ID NO: 29),DAla-DcyclohexylAla-Ala-Phe-DPhe-Nle-NH2 (SEQ ID NO: 30),DcyclohexylAla-Ala-Phe-DTrp-Lys-NH2 (SEQ ID NO: 31),DAla-DβNal-Ala-Thr-DThr-Lys-NH2 (SEQ ID NO: 32),DcyclohexylAla-Ala-Trp-DPhe-NH2 (SEQ ID NO: 33),DAla-DβNal-Ala-Ala-DAla-Lys-NH2 (SEQ ID NO: 34),DβNal-Ala-Trp-DPhe-Leu-NH2 (SEQ ID NO: 35),His-DTrp-Phe-Trp-DPhe-Lys-NH2 (SEQ ID NO: 36),DAla-DβNal-DAla-DTrp-Phe-Lys-NH2 (SEQ ID NO: 37),βAla-Trp-DAla-DTrp-Phe-NH2 (SEQ ID NO: 38),His-Trp-DAla-DTrp-Phe-Lys-NH2 (SEQ ID NO: 39),DLys-DβNal-Ala-Trp-DPhe-Lys-NH2 (SEQ ID NO: 40),DAla-DβNal-DLys-DTrp-Phe-Lys-NH2 (SEQ ID NO: 41), Tyr-DAla-Phe-Aib-NH2(SEQ ID NO: 42), Tyr-DAla-Sar-NMePhe-NH2 (SEQ ID NO: 43),αγAbu-DTrp-DTrp-Ser-NH2 (SEQ ID NO: 44), αγAbu-DTrp-DTrp-Lys-NH2 (SEQ IDNO: 45), αγAbu-DTrp-DTrp-Orn-NH2 (SEQ ID NO: 46), αAbu-DTrp-DTrp-Orn-NH2(SEQ ID NO: 47), DThr-DαNal-DTrp-DPro-Arg-NH2 (SEQ ID NO: 48),DAla-Ala-DAla-DTrp-Phe-Lys-NH2 (SEQ ID NO: 49),Alaψ[CH2NH]His-DTrp-Ala-Trp-DPhe-Lys-NH2 (SEQ ID NO: 50),Lys-DHis-DTrp-Phe-NH2 (SEQ ID NO: 51), γAbu-DTrp-DTrp-Orn-NH2 (SEQ IDNO: 52), inip-Trp-Trp-Phe-NH2 (SEQ ID NO: 53), Ac-DTrp-Phe-DTrp-Leu-NH2(SEQ ID NO: 54), Ac-DTrp-Phe-DTrp-Lys-NH2 (SEQ ID NO: 55),Ac-DTrp-DTrp-Lys-NH2 (SEQ ID NO: 56), DLys-Tyr-DTrp-DTrp-Phe-Lys-NH2(SEQ ID NO: 57), Ac-DβNal-Leu-Pro-NH2 (SEQ ID NO: 58),βAla-Trp-DTrp-DTrp-Orn-NH2 (SEQ ID NO: 59), DVal-DαNal-DTrp-Phe-Arg-NH2(SEQ ID NO: 60), DLeu-DαNal-DTrp-Phe-Arg-NH2 (SEQ ID NO: 61),CyclohexylAla-DαNal-DTrp-Phe-Arg-NH2 (SEQ ID NO: 62),DTrp-DαNal-DTrp-Phe-Arg-NH2 (SEQ ID NO: 63), DAla-DβNal-DPro-Phe-Arg-NH2(SEQ ID NO: 64), Ac-DαNal-DTrp-Phe-Arg-NH2 (SEQ ID NO: 65),DαNal-DTrp-Phe-Arg-NH2 (SEQ ID NO: 66), His-DTrp-DTrp-Lys-NH2 (SEQ IDNO: 67), Ac-DβNal-DTrp-NH2 (SEQ ID NO: 68), αAib-DTrp-DcyclohexylAla-NH2(SEQ ID NO: 69), αAib-DTrp-DAla-cyclohexylAla-NH2 (SEQ ID NO: 70),DAla-DcyclohexylAla-Ala-Ala-Phe-DPhe-Nle-NH2 (SEQ ID NO: 71),DPhe-Ala-Phe-DPal-NH2 (SEQ ID NO: 72), DPhe-Ala-Phe-DPhe-Lys-NH2 (SEQ IDNO: 73), DLys-Tyr-DTrp-DTrp-Phe-NH2 (SEQ ID NO: 74),Ac-DLys-Tyr-DTrp-DTrp-Phe-NH2 (SEQ ID NO: 75),Arg-DTrp-Leu-Tyr-Trp-Pro(cyclic Arg-Pro) (SEQ ID NO: 76),Ac-DβNal-PicLys-ILys-DPhe-NH2 (SEQ ID NO: 77), DPal-Phe-DTrp-Phe-Met-NH2(SEQ ID NO: 78), DPhe-Trp-DPhe-Phe-Met-NH2 (SEQ ID NO: 79),DPal-Trp-DPhe-Phe-Met-NH2 (SEQ ID NO: 80), βAla-Pal-DTrp-DTrp-Orn-NH2(SEQ ID NO: 81), αγAbu-Trp-DTrp-DTrp-Orn-NH2 (SEQ ID NO: 82),βAla-Trp-DTrp-DTrp-Lys-NH2 (SEQ ID NO: 83), γAbu-Trp-DTrp-DTrp-Orn-NH2(SEQ ID NO: 84), Ava-Trp-DTrp-DTrp-Orn-NH2 (SEQ ID NO: 85),DLys-Tyr-DTrp-Ala-Trp-DPhe-NH2 (SEQ ID NO: 86),His-DTrp-DArg-Trp-DPhe-NH2 (SEQ ID NO: 87), <Glu-His-Trp-DSer-DArg-NH2(SEQ ID NO: 88), DPhe-DPhe-DTrp-Met-DLys-NH2 (SEQ ID NO: 89),O-(2-methylallyl)benzophonone oxime,(R)-2-amino-3-(1H-indol-3-yl)-1-(4-phenylpiperidin-1-yl)propan-1-one,N—((R)-1-((R)-1-((S)-3-(1H-indol-3-yl)-1-oxo-1-(4-phenylpiperidin-1-yl)propan-2-ylamino)-6-amino-1-oxohexan-2-ylamino)-3-hydroxy-1-oxopropan-2-yl)benzamide,(S)—N—((S)-3-(1H-indol-3-yl)-1-oxo-1-(4-phenylpiperidin-1-yl)propan-2-yl)-6-acetamido-2-((S)-2-amino-3-(benzyloxy)propanamido)hexanamide,(S)—N—((R)-3-(1H-indol-3-yl)-1-oxo-1-(4-phenylpiperidin-1-yl)propan-2-yl)-2-((S)-2-acetamido-3-(benzyloxy)propanamido)-6-aminohexanamide,(R)—N-β-(1H-indol-3-yl)-1-(4-(2-methoxyphenyl)piperidin-1-yl)-1-oxopropan-2-yl)-4-aminobutanamide,(R)—N-(3-(1H-indol-3-yl)-1-(4-(2-methoxyphenyl)piperidin-1-yl)-1-oxopropan-2-yl)-2-amino-2-methylpropanamide,methyl 3-(p-tolylcarbamoyl)-2-naphthoate, ethyl3-(4-(2-methoxyphenyl)piperidine-1-carbonyl)-2-naphthoate,3-(2-methoxyphenylcarbamoyl)-2-naphthoate,(S)-2,4-diamino-N—((R)-3-(naphthalen-2-ylmethoxy)-1-oxo-1-(4-phenylpiperidin-1-yl)propan-2-yl)butanamide,naphthalene-2,3-diylbis((4-(2-methoxyphenyl)piperazin-1-yl)methanone),(R)-2-amino-N-β-(benzyloxy)-1-oxo-1-(4-phenylpiperazin-1-yl)propan-2-yl)-2-methylpropanamide,or (R)-2-amino-3-(benzyloxy)-1-(4-phenylpiperazin-1-yl)propan-1-one, andpharmaceutically acceptable salts, prodrugs, or active metabolitesthereof.

The antagonists embodied in the invention can be synthesized accordingto the usual methods of solution and solid phase peptide chemistry, orby classical methods known in the art. Purification of peptides is wellknown in the art and can be, for example, HPLC. Methods describinguseful peptide synthesis and purification methods can be found, forexample, in U.S. Patent Application No. 20060084607.

Synthesis of Peptides

Peptides described herein can be synthetically constructed by suitableknown peptide polymerization techniques, such as exclusively solid phasetechniques, partial solid-phase techniques, fragment condensation orclassical solution couplings. For example, the peptides of the inventioncan be synthesized by the solid phase method using standard methodsbased on either t-butyloxycarbonyl (BOC) or 9-fluorenylmethoxy-carbonyl(FMOC) protecting groups. This methodology is described by G. B. Fieldset al. in Synthetic Peptides: A User's Guide, W. M. Freeman & Company,New York, N.Y., pp. 77-183 (1992) and in the textbook “Solid-PhaseSynthesis”, Stewart & Young, Freemen & Company, San Francisco, 1969, andare exemplified by the disclosure of U.S. Pat. No. 4,105,603, issuedAug. 8, 1979. Classical solution synthesis is described in detail in“Methoden der Organischen Chemic (Houben-Weyl): Synthese von Peptiden”,E. Wunsch (editor) (1974) Georg Thieme Verlag, Stuttgart West Germany.The fragment condensation method of synthesis is exemplified in U.S.Pat. No. 3,972,859. Other available syntheses are exemplified in U.S.Pat. No. 3,842,067, U.S. Pat. No. 3,872,925, issued Jan. 28, 1975,Merrifield B, Protein Science (1996), 5: 1947-1951; The chemicalsynthesis of proteins; Mutter M, Int J Pept Protein Res 1979 March; 13(3): 274-7 Studies on the coupling rates in liquid-phase peptidesynthesis using competition experiments; and Solid Phase PeptideSynthesis in the series Methods in Enzymology (Fields, G. B. (1997)Solid-Phase Peptide Synthesis. Academic Press, San Diego. #9830).Contents of all of the foregoing disclosures are incorporated herein byreference.

Methods for preparing peptide mimetics include modifying the N-terminalamino group, the C-terminal carboxyl group, and/or changing one or moreof the amino linkages in the peptide to a non-amino linkage. Two or moresuch modifications can be coupled in one peptide mimetic inhibitor. Thefollowing are examples of modifications of peptides to produce peptidemimetics as described in U.S. Pat. Nos. 5,643,873 and 5,654,276, thecontent of both of which is incorporated herein by reference.

Designing Peptide Mimetics

Methods of designing peptide mimetics and screening of functionalpeptide mimetics are well known to those skilled in the art. One basicmethod of designing a molecule which mimics a known protein or peptideis first to identify the active region(s) of the known protein (forexample, in the case of an antibody-antigen interaction, one identifieswhich region(s) of the antibody that permit binding to the antigen), andthen searches for a mimetic which emulates the active region. If theactive region of a known protein is relatively small, it is anticipatedthat a mimetic will be smaller (e.g. in molecular weight) than theprotein, and correspondingly easier and cheaper to synthesize. Such amimetic could be used as a convenient substitute for the protein, as anagent for interacting with the target molecule.

For example, Reineke et al. (1999, Nature Biotechnology, 17; 271-275,contents of which is herein incorporated by reference) designed a mimicmolecule which mimics a binding site of the interleukin-10 protein usinga large library of short synthetic peptides, each of which correspondedto a short section of interleukin 10. The binding of each of thesepeptides to the target (in this case an antibody against interleukin-10)was then tested individually by an assay technique, to identifypotentially relevant peptides. Phage display libraries of peptides andalanine scanning method can be used.

Other methods for designing peptide mimetics to a particular peptide orprotein include those described in European Patent EP1206494, theSuperMimic program by Andrean Goede et. al. 2006 BMC Bioinformatics,7:11; and MIMETIC program by W. Campbell et. al., 2002, Microbiology andImmunology 46:211-215. The SuperMimic program is designed to identifycompounds that mimic parts of a protein, or positions in proteins thatare suitable for inserting mimetics. The application provides librariesthat contain peptidomimetic building blocks on the one hand and proteinstructures on the other. The search for promising peptidomimetic linkersfor a given peptide is based on the superposition of the peptide withseveral conformers of the mimetic. New synthetic elements or proteinscan be imported and used for searching. The MIMETIC computer program,which generates a series of peptides for interaction with a targetpeptide sequence is taught by W. Campbell et. al., 2002. In depthdiscussion of the topic is reviewed in “Peptide Mimetic Design with theAid of Computational Chemistry” by James R. Damewood Jr. in Reviews inComputational Chemistry Reviews in Computational Chemistry, January2007, Volume 9 Book Series: Reviews in Computational Chemistry,Editor(s): Kenny B. Lipkowitz, Donald B. BoydPrint ISBN: 9780471186397ISBN: 9780470125861 Published by John Wiley &Sons, Inc.; and in T.Tselios, et. al., Amino Acids, 14: 333-341, 1998. Content of all of thereferences described in this paragraph is herein incorporated byreference.

Methods for preparing libraries containing diverse populations ofpeptides, peptoids and peptidomimetics are well known in the art andvarious libraries are commercially available (see, for example, Eckerand Crooke, Biotechnology 13:351-360 (1995), and Blondelle et al.,Trends Anal. Chem. 14:83-92 (1995), and the references cited therein,each of which is incorporated herein by reference; see, also, Goodmanand Ro, Peptidomimetics for Drug Design, in “Burger's MedicinalChemistry and Drug Discovery” Vol. 1 (ed. M. E. Wolff; John Wiley & Sons1995), pages 803-861, and Gordon et al., J. Med. Chem. 37:1385-1401(1994), each of which is incorporated herein by reference). One skilledin the art understands that a peptide can be produced in vitro directlyor can be expressed from a nucleic acid, which can be produced in vitro.Methods of synthetic peptide and nucleic acid chemistry are well knownin the art. Content of all of the references described in this paragraphis herein incorporated by reference.

A library of peptide molecules also can be produced, for example, byconstructing a cDNA expression library from mRNA collected from a tissueof interest. Methods for producing such libraries are well known in theart (see, for example, Sambrook et. al., Molecular Cloning: A laboratorymanual (Cold Spring Harbor Laboratory Press 1989), which is incorporatedherein by reference). Preferably, a peptide encoded by the cDNA isexpressed on the surface of a cell or a virus containing the cDNA.

Administration and Formulation of GOAT Inhibitors and Ghrelin ReceptorAntagonist Compound

Thus, in connection with the administration of a ghrelin receptorantagonist compound of the invention, e. g. a compound which is“effective against” a disease or disorder indicates that administrationin a clinically appropriate manner results in a beneficial effect for atleast a statistically significant fraction of patients, such as aimprovement of symptoms, a cure, a reduction in symptoms or diseaseload, reduction in tumor mass or cell numbers, extension of life,improvement in quality of life, or other effect generally recognized aspositive by medical doctors familiar with treating the particular typeof disease or condition.

The therapeutic compositions of GOAT inhibitors and ghrelin receptorantagonist compounds can be administered intravenously (i.v.), as byinjection of a unit dose, for example. The term “unit dose” when used inreference to a therapeutic composition of the present invention refersto physically discrete units suitable as unitary dosage for the subject,each unit containing a predetermined quantity of active materialcalculated to produce the desired therapeutic effect in association withthe required diluent; i.e., carrier, or vehicle. Suitable therapeuticvehicle include, but not limited to, sterile saline, buffered phosphatesaline, lactated Ringer's saline.

The compositions are administered in a manner compatible with the dosageformulation, and in a therapeutically effective amount. The quantity tobe administered and timing depends on the subject to be treated,capacity of the subject's system to utilize the active ingredient, anddegree of therapeutic effect desired. Precise amounts of activeingredient required to be administered depend on the judgment of thepractitioner and are peculiar to each individual.

The compositions of the invention comprising GOAT inhibitors and/orghrelin receptor antagonists having the formulae I-VII can beadministered in any suitable manner, e.g., topically, parenterally, orby inhalation. The term “parenteral” includes injection, e.g., bysubcutaneous, intravenous, or intramuscular routes, also includinglocalized administration, e.g., at a site of disease or injury.Sustained release from implants is also contemplated. One skilled in thepertinent art will recognize that suitable dosages will vary, dependingupon such factors as the nature of the disorder to be treated, thepatient's body weight, age, and general condition, and the route ofadministration. Preliminary doses can be determined according to animaltests, and the scaling of dosages for human administration is performedaccording to art-accepted practices.

The amount of GOAT inhibitor and ghrelin receptor antagonist orcombination of compounds of the present invention administered will varydepending on numerous factors, e.g., the particular animal treated, itsage and sex, the desired therapeutic affect, the route of administrationand which polypeptide or combination of polypeptides are employed. Inall instances, however, a dose effective (therapeutically effectiveamount) to promote inhibition of growth hormone level in the blood ofthe recipient animal is used. The dose will depend on a combination offactors, i.e., antagonist receptor action(s), potency, efficacy,pharmacokinetics, pharmacodynamics, route of administration, method ofadministration and clinical disorder and/or metabolic status. Generally,dosage levels of between 0.0001 to 100 mg/kg of body weight daily areadministered to patients and mammals in need to obtain effectiveinhibition of growth hormone in the blood. The preferred amount canreadily be determined empirically by the skilled artisan.

In some embodiments, combinations of ghrelin receptor antagonistcompounds are used in any of the methods described herein. In otherembodiments, combinations of GOAT inhibitors are used in any of themethods described herein. When combinations of ghrelin receptorantagonist compounds and/or combinations of GOAT inhibitors are used,lower amounts of the antagonist can be used in the treatment of diseasesand disorders described herein. This occurs when one compound exhibitsynergistic effect over the activity of a second compound when used incombination.

The GOAT inhibitors and ghrelin receptor antagonist compounds can beadministrated to a subject in combination with another pharmaceuticallyactive agent or treatment modality for a particular indication.Exemplary pharmaceutically active compound include, but are not limitedto, those found in Harrison's Principles of Internal Medicine, 13^(th)Edition, Eds. T. R. Harrison et al. McGraw-Hill N.Y., NY; PhysiciansDesk Reference, 50^(th) Edition, 1997, Oradell N.J., Medical EconomicsCo.; Pharmacological Basis of Therapeutics, 8^(th) Edition, Goodman andGilman, 1990; United States Pharmacopeia, The National Formulary, USPXII NF XVII, 1990; current edition of Goodman and Oilman's ThePharmacological Basis of Therapeutics; and current edition of The MerckIndex, the complete contents of all of which are incorporated herein byreference.

In another embodiment, the GOAT inhibitors and ghrelin receptorantagonist compounds of the inventions can be used in combination withother treatment regime for treating the diseases and disordersassociated with obesity, overeating, diabetes, unregulated cellularproliferation, and cancer.

The of GOAT inhibitors and ghrelin receptor antagonist compounds of thepresent invention can be formulation for sustained or controlledrelease. The antagonists and/inhibitors of the present invention can beadmixed with biologically compatible polymers or matrices which controlthe release rate of the antagonists into the immediate environment.Controlled or sustained release compositions include formulation inlipophilic depots (e.g., fatty acids, waxes, oils). Also contemplated bythe invention are particulate compositions coated with polymers (e.g.,poloxamers or poloxamines). Other embodiments of the compositions of theinvention incorporate particulate forms, protective coatings, proteaseinhibitors or permeation enhancers for various routes of administration,including parenteral, pulmonary, nasal and oral.

Controlled release permits dosages to be administered over time, withcontrolled release kinetics. In some instances, delivery of thetherapeutic agent is continuous to the site where treatment is needed,for example, over several weeks. Controlled release over time, forexample, over several days or weeks, or longer, permits continuousdelivery of the therapeutic agent to obtain optimal treatment. Thecontrolled delivery vehicle is advantageous because it protects thetherapeutic agent from degradation in vivo in body fluids and tissue,for example, by proteases.

Controlled release from the pharmaceutical formulation may be designedto occur over time, for example, for greater than about 12 or 24 hours.The time of release may be selected, for example, to occur over a timeperiod of about 12 hours to 24 hours; about 12 hours to 42 hours; or, e.g., about 12 to 72 hours. In another embodiment, release may occur forexample on the order of about 2 to 90 days, for example, about 3 to 60days. In one embodiment, the GOAT inhibitor and/or ghrelin receptorantagonist is delivered locally over a time period of about 7-21 days,or about 3 to 10 days. In other instances, the therapeutic agent isadministered over 1,2,3 or more weeks in a controlled dosage. Thecontrolled release time may be selected based on the condition treated.

“Controlled release” also encompasses patterned delivery (e.g.,intermittent delivery over a period of time that is interrupted byregular or irregular time intervals). Without limitation, theintermittent delivery can be interrupted by a period of at least 30minutes, at least 1 hour, at least 2 hours, at least 3 hours, at least 4hours, at least 5 hours, at least 6 hours, at least 12, at least 24hours, at least 1 day, at least 2 days, at least 3 days, at least 4days, at least 5 days, at least 6 days, at least 1 week, at least onemonth or more

As used herein, the terms “pharmaceutically acceptable”,“physiologically tolerable” and grammatical variations thereof, as theyrefer to compositions, carriers, diluents and reagents, are usedinterchangeably and represent that the materials are capable ofadministration to or upon a mammal without the production of undesirablephysiological effects.

Compositions comprising an effective amount of a ghrelin receptorantagonist and GOAT inhibitor, in combination with other components,such as a physiologically acceptable diluent, carrier, or excipient, areprovided herein. The antagonists/inhibitor can be formulated accordingto known methods used to prepare pharmaceutically useful compositions.They can be combined in admixture, either as the sole active material orwith other known materials suitable for a given indication, withpharmaceutically acceptable diluents (e.g., saline, Tris-HCl, acetate,and phosphate buffered solutions), preservatives (e.g., thimerosal,benzyl alcohol, parabens), emulsifiers, solubilizers, adjuvants and/orcarriers. Suitable formulations for pharmaceutical compositions includethose described in Remington's Pharmaceutical Sciences, 16^(th) ed.1980, Mack Publishing Company, Easton, Pa., content of which is hereinincorporated by reference.

In addition, such compositions can be complexed with polyethylene glycol(PEG), metal ions, or incorporated into polymeric compounds such aspolyacetic acid, polyglycolic acid, hydrogels, dextran, etc., orincorporated into liposomes, microemulsions, micelles, unilamellar ormultilamellar vesicles, erythrocyte ghosts or spheroblasts. Suchcompositions will influence the physical state, solubility, stability,rate of in vivo release, and rate of in vivo clearance, and are thuschosen according to the intended application.

Toxicity and therapeutic efficacy can be determined by standardpharmaceutical procedures in cell cultures or experimental animals,e.g., for determining the LD50 (the dose lethal to 50% of thepopulation) and the ED50 (the dose therapeutically effective in 50% ofthe population). The dose ratio between toxic and therapeutic effects isthe therapeutic index and it can be expressed as the ratio D50/ED50.Compositions that exhibit large therapeutic indices, are preferred.

The data obtained from the cell culture assays and animal studies can beused in formulating a range of dosage for use in humans. The dosage ofsuch compounds lies preferably within a range of circulatingconcentrations that include the ED50 with little or no toxicity. Thedosage may vary within this range depending upon the dosage formemployed and the route of administration utilized.

The therapeutically effective dose can be estimated initially from cellculture assays. A dose may be formulated in animal models to achieve acirculating plasma concentration range that includes the IC50 (i.e., theconcentration of the therapeutic which achieves a half-maximalinhibition of symptoms) as determined in cell culture. Levels in plasmamay be measured, for example, by high performance liquid chromatography.The effects of any particular dosage can be monitored by a suitablebioassay.

The dosage may be determined by a physician and adjusted, as necessary,to suit observed effects of the treatment. Generally, the compositionsare administered so that a GOAT inhibitor and/or ghrelin receptorantagonist is given at a dose from 1 μg/kg to 150 mg/kg, 1 μg/kg to 100mg/kg, 1 μg/kg to 50 mg/kg, 1 μg/kg to 20 mg/kg, 1 μg/kg to 10 mg/kg, 1μg/kg to 1 mg/kg, 100 μg/kg to 100 mg/kg, 100 μg/kg to 50 mg/kg, 100μg/kg to 20 mg/kg, 100 μg/kg to 10 mg/kg, 100 μg/kg to 1 mg/kg, 1 mg/kgto 100 mg/kg, 1 mg/kg to 50 mg/kg, 1 mg/kg to 20 mg/kg, 1 mg/kg to 10mg/kg, 10 mg/kg to 100 mg/kg, 10 mg/kg to 50 mg/kg, or 10 mg/kg to 20mg/kg. It is to be understood that ranges given here include allintermediate ranges, for example, the range 1 mg/kg to 10 mg/kg includes1 mg/kg to 2 mg/kg, 1 mg/kg to 3 mg/kg, 1 mg/kg to 4 mg/kg, 1 mg/kg to 5mg/kg, 1 mg/kg to 6 mg/kg, 1 mg/kg to 7 mg/kg, 1 mg/kg to 8 mg/kg, 1mg/kg to 9 mg/kg, 2 mg/kg to 10 mg/kg, 3 mg/kg to 10 mg/kg, 4 mg/kg to10 mg/kg, 5 mg/kg to 10 mg/kg, 6 mg/kg to 10 mg/kg, 7 mg/kg to 10 mg/kg,8 mg/kg to 10 mg/kg, 9 mg/kg to 10 mg/kg, and the like. It is to befurther understood that the ranges intermediate to the given above arealso within the scope of this invention, for example, in the range 1mg/kg to 10 mg/kg, dose ranges such as 2 mg/kg to 8 mg/kg, 3 mg/kg to 7mg/kg, 4 mg/kg to 6 mg/kg, and the like.

With respect to duration and frequency of treatment, it is typical forskilled clinicians to monitor subjects in order to determine when thetreatment is providing therapeutic benefit, and to determine whether toincrease or decrease dosage, increase or decrease administrationfrequency, discontinue treatment, resume treatment or make otheralteration to treatment regimen. The dosing schedule can vary from oncea week to daily depending on a number of clinical factors, such as thesubject's sensitivity to the polypeptides. The desired dose can beadministered at one time or divided into subdoses, e.g., 2-4 subdosesand administered over a period of time, e.g., at appropriate intervalsthrough the day or other appropriate schedule. Such sub-doses can beadministered as unit dosage forms. In some embodiments, administrationis chronic, e.g., one or more doses daily over a period of weeks ormonths. Examples of dosing schedules are administration daily, twicedaily, three times daily or four or more times daily over a period of 1week, 2 weeks, 3 weeks, 4 weeks, 1 month, 2 months, 3 months, 4 months,5 months, or 6 months or more.

Treatment of Diseases and Disorders

By “treatment”, “prevention” or “amelioration” of a disease or disorderis meant delaying or preventing the onset of such a disease or disorder,reversing, alleviating, ameliorating, inhibiting, slowing down orstopping the progression, aggravation, deterioration or severity of acondition associated with such a disease or disorder. In one embodiment,the symptoms of a disease or disorder are alleviated by at least 5%, atleast 10%, at least 20%, at least 30%, at least 40%, or at least 50%.The term “treatment” is intended to encompass also prophylaxis, therapyand cure.

In one embodiment, a method to treat obesity is encompassed. Inparticular, the present invention comprises methods for regulating foodintake in a human subject; for improving a compliance of a human subjectto caloric restriction; and for reducing a desire of a human subject toconsume an over-abundance of calories and/or fats. This method comprisesthe administration of an effective amount of ghrelin receptor antagonistand/or an effective amount of ghrelin O-acyltransferase (GOAT) inhibitoras described above.

In some embodiments, the food intake is reduced by at least 5%, at least10%, at least 20%, at least 30%, at least 40%, at least 50%, at least60%, at least 70%, at least 80%, at least 90%, at least 95% or morerelative to subject that is not undergoing treatment. The amount of foodintake can be based on the number of calories or on the weight of food.

The present invention further provides a method for preventing orreducing weight gain in a human subject, by administration of aneffective amount of ghrelin receptor antagonist and/or an effectiveamount of GOAT inhibitor that have a pharmacological half-life thatallows an efficient treatment regime thereof.

Also encompassed are methods for reducing a desire of a human subject toconsume calories following gastric banding or gastric bypass surgery, byadministration of an effective amount of a GOAT inhibitor and/or aneffective amount of a ghrelin receptor antagonist.

In addition to the obesity related disorders discussed above,simultaneous administration of ghrelin receptor antagonist compositionsand GOAT inhibitors are useful in the treatment or prevention of thefollowing obesity related diseases and/or disorders: overeating;bulimia; hypertension; diabetes, elevated plasma insulin concentrations;insulin resistance; dyslipidemias; hyperlipidemia; endometrial, breast,prostate and colon cancer; osteoarthritis; obstructive sleep apnea;cholelithiasis; gallstones; abnormal heart rhythms; heart arrythymias;myocardial infarction; congestive heart failure; coronary heart disease;sudden death; stroke; polycystic ovarian disease; craniopharyngioma; thePrader-Willi Syndrome; Frohlich's syndrome; GH-deficient subjects;normal variant short stature; Turner's syndrome; and other pathologicalconditions showing reduced metabolic activity or a decrease in restingenergy expenditure as a percentage of total fat-free mass, e. g.,children with acute lymphoblastic leukemia.

The present invention also comprise treating of obesity and obesityrelated diseases and disorders by administering a combination of aghrelin receptor antagonist and a GOAT inhibitor together with ananti-obesity agent, which can be administered separately orconcurrently. In other embodiments, several ghrelin receptor antagonistsand GOAT inhibitors are used.

Anti-obesity agents to be used in combination with the gherlin receptorantagonists and GOAT inhibitors of the present invention are known tothose of skill in the art and can include, but not limited to, a 5HT(serotonin) transporter inhibitor, a NE (norepinephrine) transporterinhibitor, a CB-1 (cannabinoid-1) antagonist/inverse agonist, a H3(histamine H3) antagonist/inverse agonist, a MCH1R (melaninconcentrating hormone 1R) antagonist, a MCH2R agonist/antagonist, a NPY1antagonist, a leptin, a leptin derivative, a leptin analog, an opioidantagonist, an orexin antagonist, a BRS3 (bombesin receptor subtype 3)agonist, a CCK-A (cholecystokinin-A) agonist, a CNTF (Ciliaryneurotrophic factor), a CNTF derivative, a GHS (growth hormonesecretagogue receptor) agonist, a 5HT2C (serotonin receptor 2C) agonist,a Mc4r (melanocortin 4 receptor) agonist, a monoamine reuptakeinhibitor, an UCP-1 (uncoupling protein-1), 2, or 3 activator, a β3(beta adrenergic receptor 3) agonist, a thyroid hormone β agonist, a PDE(phosphodiesterase) inhibitor, a FAS (fatty acid synthase) inhibitor, aDGAT1 (diacylglycerol acyltransferase) inhibitor, a DGAT2 inhibitor, anACC2 (acetyl-CoA carboxylase-2) inhibitor, a glucocorticoid antagonist,an acyl-estrogen, a lipase inhibitor, a fatty acid transporterinhibitor, a dicarboxylate transporter inhibitor, a glucose transporterinhibitor, a serotonin reuptake inhibitors, metformin, and topiramate.

The anti-obesity compound to be used in combination with the ghrelinreceptor antagonists of the present invention may act via a mechanismother than ghrelin, thus providing for additive anti-obesity effects.

In one embodiment, the ghrelin receptor antagonists and GOAT inhibitorsof the present invention is administered prior to taking a meal, forexample, 4 hours, 3 hours, 2 hours, 1 hour, or 0.5 hours prior toexpected meal time. Preferably, the ghrelin receptor antagonist and GOATinhibitor are administered 0.5 hours prior to feeding. Alternatively,the ghrelin receptor antagonist and GOAT inhibitor can be administeredcontinuously, for example, systemically, as a single administrationevery 6, 5, 4, 3, 2, or 1 month, preferably every 3 months. Here, theghrelin receptor antagonist and GOAT inhibitor of the present inventioncan normalize an otherwise dysfunctional endocrine system. The compoundcan be active in the individual for several months.

In another embodiment of the present invention, a method for treatingdiabetes is encompassed. Diabetes can include both insulin-dependentdiabetes mellitus (i.e., IDDM, also known as type I diabetes) andnon-insulin-dependent diabetes mellitus (i.e., NIDDM, also known as TypeII diabetes). In this method, individuals with or at risk for developingdiabetes are administered the ghrelin receptor antagonists and GOATinhibitors of the present invention alone or in combination with otherdiabetes treatments known to those of skill in the art.

“Treatment” (of obesity and obesity-related disorders) refers to theadministration of the compounds or combinations of the present inventionto reduce or maintain the body weight of an obese subject. One outcomeof treatment can be reducing the body weight of an obese subjectrelative to that subject's body weight immediately before theadministration of the compounds or combinations of the presentinvention. Another outcome of treatment can be preventing body weightregain of body weight previously lost as a result of diet, exercise, orpharmacotherapy. Another outcome of treatment can be decreasing theoccurrence of and/or the severity of obesity-related diseases. Anotheroutcome of treatment may be to maintain weight loss. The treatment cansuitably result in a reduction in food or calorie intake by the subject,including a reduction in total food intake, or a reduction of intake ofspecific components of the diet such as carbohydrates or fats; and/orthe inhibition of nutrient absorption; and/or the inhibition of thereduction of metabolic rate; and in weight reduction in patients in needthereof. The treatment can also result in an alteration of metabolicrate, such as an increase in metabolic rate, rather than or in additionto an inhibition of the reduction of metabolic rate; and/or inminimization of the metabolic resistance that normally results fromweight loss.

“Treatment” (of cancer) refers to the administration of the compounds orcombinations of the present invention to reduce, eliminate, amelioratethe symptoms brought about directly or indirectly by the cancer, e. g.the symptoms of Cushing disease as a result of excess ACTH from apituitary tumor or cctopic acromegaly from excess growth hormones from ametastatic bronchial carcinoid tumor.

In one embodiment, the ghrelin receptor antagonists and GOAT inhibitorsof the present invention are used to treat or prevent hormonallyfunctional endocrine or non-endocrine tumors.

In one embodiment, the combined used of the ghrelin receptor antagonistsand GOAT inhibitors described herein provides a method to decrease orregulate gastrointestinal motility or acidity in a mammal.

In another embodiment, the combined used of the ghrelin receptorantagonists and GOAT inhibitors described herein provide a method oftreatment, prevention or management of psychobehavior related to underand over-nutrition such as hunger, satiety and anxiety.

In yet another embodiment, the combined used of the ghrelin receptorantagonists and GOAT inhibitors described herein provides a method foraugmenting the actions of desacyl ghrelin by decreasing the action ofacyl ghrelin and its receptor.

In another embodiment, the ghrelin receptor antagonists and GOATinhibitors described herein are administered, in combination, to asubject for the treatment of pituitary tumor, e. g. to inhibit pituitarytumor producing growth hormone.

In another embodiment, the ghrelin receptor antagonists and GOATinhibitors described herein are administered in conjunction with methodsfor the treatment of tumors that produce prolactin. Prolactin (“PRL”) isa 23-kDa neuroendocrine hormone which is structurally related to growthhormone. Prolactin secretion has been associated with several types ofcancer including, but not limited to breast, pituitary and prostrate.Thus, the present invention relates to methods for inhibiting the cellproliferation-promoting effects of prolactin on its receptor. Conditionswhich can benefit from the administration of both a ghrelin receptorantagonist and a GOAT inhibitor include both benign and malignantproliferation of cells which express a prolactin receptor. Suchconditions include but are not limited to proliferative diseases of thebreast, including benign conditions such as breast adenomas andfibrocystic disease, and malignant conditions such as breast cancer,including ductal, scirrhous, medullary, colloid and lobular carcinomas(local or metastatic); and proliferative diseases of the prostate,including benign prostatic hypertrophy and prostate cancer (local ormetastatic).

Also encompassed are methods for the treatment of metabolic syndrome.The term “metabolic syndrome”, also known as syndrome X, is defined inthe Third Report of the National Cholesterol Education Program ExpertPanel on Detection, Evaluation and Treatment of High Blood Cholesterolin Adults (ATP-III). E. S. Ford et al., JAMA, vol. 287 (3), Jan. 16,2002, pp 356-359. Briefly, a person is defined as having metabolicsyndrome if the person has three or more of the following symptoms:abdominal obesity, hypertriglyceridemia, low HDL cholesterol, high bloodpressure, and high fasting plasma glucose. The criteria for these aredefined in ATP-III. Metabolic syndrome is obesity related and thusadministration of the novel ghrelin receptor antagonists of the presentinvention are useful in its treatment.

Furthermore, methods for the diagnosis of obesity and obesity relateddiseases and disorders, including diabetes are encompassed. In thisembodiment, the ghrelin receptor antagonist(s) and GOAT inhibitor(s) areadministered to a subject and their response is closely analyzed. Adecrease in desire for food immediately following administration of thecompounds or a decrease in weight gain or a reduction in weightindicates a propensity to or a current affliction with obesity or anobesity related disease or disorder.

In one embodiment, Ghrelin/GHRP/GHS receptor antagonists can be utilizedas a diagnostic agent to assess the role of ghrelin, other ghrelin-likemolecules, and ghrelin receptor agonists or its receptor in theregulation of GH secretion, food intake, and gastrointestinal motility.The antagonists can also be used to rule out endogenouspathophysiological activities of ghrelin, assess the role of ghrelin invarious physiological and metabolic processes, assess the effects ofexogenous ghrelin, GHSs as well as other agents that possibly act viarelease of endogenous ghrelin or via ghrelin mimics, and determinebiological actions of acylated ghrelin over that of desacylated ghrelin.

Specifically, in one embodiment diagnostic indicates/reveals a role ofghrelin and/or its receptor(s) in over, under or dysfunctional secretionin the pathophysiology of GH release. In another embodiment, diagnosticindicates/reveals a role of ghrelin and/or its receptor(s) in thepathophysiology of food intake in over or under nutrition. In yetanother embodiment, diagnostic indicates/reveals risk of developingobesity, metabolic syndrome, diabetes and/or success rate ofanti-obesity therapy. In yet another embodiment, diagnosticindicates/reveals a role of ghrelin and/or its receptor(s) incognitive-memory and psychobehavior related to under and/or or nutritionsuch as hunger, satiety and anxiety. In another embodiment, thediagnostic indicates/reveals role of ghrelin and/or its receptor(s) inthe pathophysiology of insulin secretion and/or its actions. In anotherembodiment, the diagnostic indicates/reveals role of ghrelin and itsreceptor(s) on the pathophysiology of gastrointestinal (GI) motility,acidity or other GI disorders. In yet another embodiment, the diagnosticmethod indicates/reveals role of ghrelin and its receptor(s) onendothelial dysfunction in particular related to vasoconstriction and/orinsulin actions in particular hypertension, diabetes and metabolicsyndrome. In another embodiment, the diagnostic method indicates/revealsrole and action of ghrelin and its receptor(s) on hepaticgluconeogenesis and body fat as indicated by effects on circulatingglucose, insulin, adipokines, leptin, resistin, adiponectin andplasminogen activator inhibitor. In still another embodiment, thediagnostic indicates/reveals/distinguishs the actions of acylatedghrelin and desacylated ghrelin on selective actions of certain GHsecretagogues. In another method, the diagnostic methodindicates/reveals a role of ghrelin and its receptor(s) on agents thatincrease (i.e., anti-depressants, glucocorticoids and other drugs thatinfluence food intake). In still another embodiment, the diagnosticmethod indicates/reveals a role of ghrelin and its receptor(s) inhormonally functional endocrine and non-endocrine tumors.

In another embodiment, the ghrelin receptor antagonists and GOATinhibitors described herein are administered separately or concurrentlyto a subject in need thereof.

In one embodiment, provided herein is a method for treatment, preventionor management of obesity in a subject, the method comprising the step ofadministering an effective amount of a compound having the formula I,wherein the compound is a ghrelin receptor antagonist.

In another embodiment, the method of for treatment, prevention ormanagement of obesity in a subject further comprises an anti-obesitytreatment. In another embodiment, the anti-obesity treatment is acombination of dietary restriction therapy with a 5HT (serotonin)transporter inhibitor, a NE (norepinephrine) transporter inhibitor, aCB-1 (cannabinoid-1) antagonist/inverse agonist, a H3 (histamine H3)antagonist/inverse agonist, a MCH1R (melanin concentrating hormone 1R)antagonist, a MCH2R agonist/antagonist, a NPY1 antagonist, leptin, aleptin derivative, a leptin analog, an opioid antagonist, an orexinantagonist, a BRS3 (bombesin receptor subtype 3) agonist, a CCK-A(cholecystokinin-A) agonist, a CNTF (Ciliary neurotrophic factor), aCNTF derivative, a lipase drug inhibitor, an inhibitor of food intake,an incretin, an incretin agonist, an incretin analog or an incretinmimic administered simultaneously, concurrently or sequentially.

In one embodiment, provided herein is a method for treatment, preventionor management of diabetes mellitus in a subject in need thereof, themethod comprising the step of administering an effective amount of acompound having the formulae (I)-(VII), wherein the compound is aghrelin receptor antagonist.

In one embodiment, provided herein is a method for the modulation ofghrelin receptor in a subject in need thereof, the method comprising thestep of administering an effective amount of a GOAT inhibitor.

In one embodiment, provided herein is a method for the modulation ofghrelin receptor in a subject in need thereof, the method comprising thestep of administering an effective amount of a compound having theformulae (I)-(VII), wherein the compound is a ghrelin receptorantagonist.

In one embodiment, provided herein is a method for treatment,prevention, or management of metabolic syndrome in a subject in needthereof, the method comprising the step of administering an effectiveamount of a compound having the formulae (I)-(VII), wherein the compoundis a ghrelin receptor antagonist.

In one embodiment, provided herein is a method for treatment,prevention, or management of cancer in a subject in need thereof, themethod comprising the step of administering an effective amount of acompound having the formulae (I)-(VII), wherein the compound is aghrelin receptor antagonist.

The present invention can be defined in any of the following numberedparagraphs:

-   1. A method for treatment, prevention or management of obesity or    obesity related disease or disorder in a subject in need thereof,    said method comprising the step of administering an effective amount    of a ghrelin O-acyltransferase (GOAT) inhibitor and an effective    amount of a ghrelin receptor antagonist.-   2. A method for treatment, prevention or management of obesity or    obesity related disease or disorder in a subject in need thereof,    said method comprising the step of administering an effective amount    of a ghrelin O-acyltransferase (GOAT) inhibitor.-   3. The method of paragraph 1 or 2, further comprising an    anti-obesity treatment.-   4. The method of paragraph 3, wherein the anti-obesity treatment is    a combination of dietary restriction therapy with a 5HT (serotonin)    transporter inhibitor, a NE (norepinephrine) transporter inhibitor,    a CB-1 (cannabinoid-1) antagonist/inverse agonist, a H3 (histamine    H3) antagonist/inverse agonist, a MCH1R (melanin concentrating    hormone 1R) antagonist, a MCH2R agonist/antagonist, a NPY1    antagonist, a leptin, a leptin derivative, a leptin analog,    PYY(1-36), PYY(3-36), an opioid antagonist, an orexin antagonist, a    BRS3 (bombesin receptor subtype 3) agonist, a CCK-A    (cholecystokinin-A) agonist, a CNTF (Ciliary neurotrophic factor), a    CNTF derivative, a lipase drug inhibitor, an inhibitor of food    intake, an incretin, an incretin agonist, an incretin analog or an    incretin mimic administered simultaneously, concurrently or    sequentially.-   5. The method of paragraph 4, wherein the inhibitor of food intake    is glucagon-like-peptide (Glip-1/Glip-2) or oxyntomodulin or their    analogs, derivatives, or mimics.-   6. The method of paragraph 3, wherein the anti-obesity treatment is    bariatric surgery.-   7. A method for the modulation of ghrelin receptor in a subject in    need thereof, said method comprising the step of administering an    effective amount of a ghrelin receptor antagonist.-   8. The method paragraph 7, further comprising administering an    effective amount of a GOAT inhibitor.-   9. A method for treatment, prevention or management of diabetes    mellitus in a subject in need thereof, said method comprising the    step of administering an effective amount of a GOAT inhibitor and/or    an effective amount of a ghrelin receptor antagonist.-   10. The method of paragraph 9, wherein the diabetes mellitus is type    I or II.-   11. A method for treatment, prevention, or management of metabolic    syndrome in a subject in need thereof, said method comprising the    step of administering an effective amount of a GOAT inhibitor and/or    an effective amount of a ghrelin receptor antagonist.-   12. The method of any of paragraphs 9-11, wherein the ghrelin    receptor antagonist is administered alone, sequentially, or    concomitantly with a biguanide, a peroxisome proliferator    activator-receptor alpha (PPAR-alpha) ligand or PPAR-gamma ligand.-   13. A method for treatment, prevention, or management of cancer in a    subject in need thereof, said method comprising the step of    administering an effective amount of a GOAT inhibitor and/or an    effective amount of a ghrelin receptor antagonist.-   14. The method of any of paragraph 1-13, wherein the subject is    obese or at risk of obesity.-   15. The method of any of paragraphs 1-14, wherein the subject has a    body mass index (BMI) of over 25.-   16. The method of any of paragraphs 1-15, wherein the subject has a    body mass index (BMI) of between 25 and 30.-   17. The method of any of paragraphs 1-16, wherein the subject has a    body mass index (BMI) of over 30.-   18. The method of any of paragraphs 1-17, wherein GH secretion in    the subject is inhibited by at least at least 5%, at least 10%, at    least 20%, at least 30%, at least 40%, at least 50%, at least 60%,    at least 70%, at least 80%, at least 90%, at least 95% or complete    inhibition relative to GH secretion before onset of treatment or    relative to an untreated control.-   19. The method of any of paragraphs 1-18, wherein food intake by the    subject is decreased or inhibited by at least 5%, at least 10%, at    least 20%, at least 30%, at least 40%, at least 50%, at least 60%,    at least 70%, at least 80%, at least 90%, at least 95% or complete    inhibition relative to an untreated control.-   20. The method of any of paragraphs 1-19, wherein the GOAT inhibitor    and/or the ghrelin receptor antagonist comprises an octanoylated    peptide and wherein the octanoylation is at position three of the    peptide.-   21. The method of any of paragraphs 1-19, wherein the GOAT inhibitor    and/or the ghrelin receptor antagonist comprises an octanoylated    pentapeptide and wherein the octanoylation is at position three of    the pentapeptide.-   22. The method of any of paragraphs 1-20, wherein the GOAT inhibitor    and/or the ghrelin receptor antagonist comprises an octanoylated    pentapeptide and wherein the octanoylation is on the side chain of    residue at position three of the pentapeptide-   23. The method of any of paragraphs 1-21, wherein the octanoylated    ghrelin pentapeptide is Gly-Ser-[Ser-Octanoyl³]-Phe-LeuCO₂H (SEQ ID    NO: 1), Gly-Ser-[Ser-Octanoyl³]-Phe-LeuNH₂ (SEQ ID NO: 94),    Gly-Ser-[Ser-Octanoyl³]-Phe-Leu-CO₂H (SEQ ID NO: 1),    Gly-Ser-[Dap-Octanoyl³]-Phe-LeuCO₂H (SEQ ID NO: 2), or    Gly-Ser-[Dap-Octanoyl³]-Phe-LeuNH₂ (SEQ ID NO: 95).-   24. The method of any of paragraphs 1-22, wherein the ghrelin    receptor antagonist is of formula (I): A₁-A₂-A₃-A₄-A₅-A₆, wherein:    A₁ is His, Tyr, desamino-Tyr, D- or L-Ala, β-Ala, CyclohexylAla    (Cyclohexylalanine), D-Arg, Ava (aminovaleric acid), Gly, <Glu    (pyroglutaminic acid), α-Aib (alpha-aminoisobutyric acid), γ-Abu    (gamma-aminobutyric acid), α-Abu, α,γ-Abu, D-Val, D-phe, D-Thr,    D-Pal (pyridylalanine), D-Lys, AcD-Lys, D-Leu, D-Trp,    D,α-naphthylalanine, D,β-naphthylalanine, or Ac-D,β-naphthylalanine;    A₂ is D-α-naphthylalanine, D-β-naphthylalanine;    Ac-D-β-naphthylalanine, D- or L-Trp, D- or L-Phe, Ala, His, PicLys    (N^(ε)-picoloyl-lysine), D-Cyclohexylalanine, or an amino acid that    is methylated at the terminal nitrogen of the a carbon atom of the    A₂ residue; A₃ is D- or L-Lys, lysine analogs and derivatives, Arg,    arginine analogs and derivatives, Orn, Phe, Trp, Leu, Pro, Ala, Ser,    Pal, or α,γ-Abu; A₄ is D- or L-Trp, D- or L-Phe, Ala, Ser, Tyr, Met,    Pro, Thr, ILys, or CyclohexylAla; A₅ is D- or L-Trp, D- or L-Phe,    Ala, Lys, Arg, Orn, Thr, Leu, or D-CyclohexylAla; A₆ is Lys, Arg,    Orn, D- or L-Phe, Pro(cyclic Arg-Pro), Nle (norleucine), α,γ-Abu    amide, or a free acid carboxyl group; and prodrugs, metabolite, or    pharmaceutically acceptable salts thereof.-   25. The method of any of paragraphs 1-23, wherein the ghrelin    receptor antagonist is of formula (II): A₇-A₈-A₉-A₁₀, wherein: A₇ is    D-α-Nal, D-β-Nal, Ac-D-β-Nal, Ac-D-α-Nal, D- or L-Tyr, Ac-D-Tyr,    Lys, D-Phe, His, α-Abu, α,γ-Abu, γ-Abu, DcyclohexylAla, or    isonipecotic carboxylic acid (inip); A₈ is D- or L-Trp, Ala, His,    Phe, or Leu; A₉ is D- or L-Trp, Ala, CyclohexylAla, Phe, Pro, Lys,    Sarcosine (Sar, N-methylglycine), or a free acid carboxyl group; A₁₀    is D- or L-Arg, Phe, CyclohexylAla, Lys, Ser, NMePhe, DPal, Aib, or    Orn; and prodrugs, metabolite, or pharmaceutically acceptable salts    thereof.-   26. The method of any of paragraphs 1-23, wherein the ghrelin    receptor antagonist is of formula

wherein:

-   -   R₁ is OH; R₂ is H, —C(O)—(CH₂)₃—NH₂, —C(O)—C(CH₃)₂—NH₂,        —C(O)—CH((CH₂)₄—NHR₅)—NH—C(O)—CH(NHR₃)CH₂OR₄; R₃ is H,        —C(O)-phenyl, or —C(O)CH₃; R₄ is H, or —CH₂-phenyl; R₅ is H, or        —C(O)CH₃; and isomers, derivatives, prodrugs, metabolite, or        pharmaceutically acceptable salts thereof.

-   27. The method of any of paragraphs 1-23, wherein the ghrelin    receptor antagonist is of formula (IV):

wherein:

-   -   R₆ is —C(O)OCH₂CH₃, —C(O)OCH₃,

R₇ is —H,

and isomers, derivatives, prodrugs, metabolite, or pharmaceuticallyacceptable salts thereof.

-   28. The method of any of paragraphs 1-23, wherein the ghrelin    receptor antagonist is of formula (V):

wherein:

-   -   R₈ is H, or —C(O)C(CH₃)₂—NH₂; and isomers, derivatives,        prodrugs, metabolite, or pharmaceutically acceptable salts        thereof.

-   29. The method of any of paragraphs 1-23, wherein the ghrelin    receptor antagonist is of formula (VI): S¹B¹S²B²S³, wherein: S¹ is    H, CO₂H, R¹¹, R¹¹R¹², or R¹¹R¹²R¹³; S² is des-Amino, H, CO₂H, R¹¹,    or R¹¹R¹², or R¹¹R¹²R₁₃; S³ is H, CO₂H, NH₂, R¹¹, R¹¹R¹², or    R¹¹R¹²R¹³; R¹¹, R¹², and R¹³ are selected from the group consisting    of L-amino acids, Pal (3-pyridyl alanine), cyclo-Ala, Aib, Nle,    inip, Abu, βNal, αNal, Orn, carboxylic acid, and their respective D    isomers, and can be the same or different; B¹ is selected from the    group consisting of Trp, βNal, αNal, Leu, Lys, cyclohexylAla, and    their respective D isomers; B² is any natural L-amino acid, Pal    (3-pyridyl alanine), cycloAla, Aib, Nle, inip, Abu, βNal, αNal, Orn,    and their respective D isomers; and isomers, derivatives, prodrugs,    metabolite, or pharmaceutically acceptable salts thereof.

-   30. The method of any of paragraphs 1-23, wherein the ghrelin    receptor antagonist is of formula (VII):    A¹¹-A¹²-A¹³-Gly-Ser-A¹⁴-Phe-Leu-A¹⁵-A¹⁶-A¹⁷-A¹⁸ (SEQ ID NO: 93),    wherein each of A¹¹, A¹², and A¹³ is absent, an amino acid or an    amino protecting group, and can be the same or different; each of    A¹⁵, A¹⁶, A¹⁷, and A¹⁸ is absent or an amino acid, and can be the    same or different; A¹⁴ is a serine conjugated with a —C(O)C₁-C₂₀    alky group on the side chain OH of said serine or a diaminopropionic    acid conjugated with a —C(O)C₁-C₂₀ alky group on one of the amino    group diaminopropionic acid, provided at least one of A¹¹, A¹², or    A¹³ is present.

-   31. The method of paragraph 30, wherein the amino acid is selected    from the group consisting of alanine, arginine, asparagine, aspartic    acid, cysteine, glutamic acid, glutamine, glycine, histidine,    isoleucine, leucine, lysine, methionine, phenylalanine, proline,    serine, threonine, tryptophan, tyrosine, valine, homocysteine,    phosphoserine, phosphothreonine, phosphotyrosine, hydroxyproline,    gamma-carboxyglutamate; hippuric acid, octahydroindole-2-carboxylic    acid, statine, 1,2,3,4,-tetrahydroisoquinoline-3-carboxylic acid,    penicillamine (3-mercapto-D-valine), ornithine, citruline,    alpha-methyl-alanine, para-benzoylphenylalanine,    para-aminophenylalanine, p-fluorophenylalanine, phenylglycine,    propargylglycine, sarcosine, and tert-butylglycine), diaminobutyric    acid, 7-hydroxy-tetrahydroisoquinoline carboxylic acid,    naphthylalanine, biphenylalanine, cyclohexylalanine,    amino-isobutyric acid, norvaline, norleucine, tert-leucine,    tetrahydroisoquinoline carboxylic acid, pipecolic acid,    phenylglycine, homophenylalanine, cyclohexylglycine, dehydroleucine,    2,2-diethylglycine, 1-amino-1-cyclopentanecarboxylic acid,    1-amino-1-cyclohexanecarboxylic acid, amino-benzoic acid,    amino-naphthoic acid, gamma-aminobutyric acid,    difluorophenylalanine, nipecotic acid, alpha-amino butyric acid,    thienyl-alanine, t-butylglycine, desamino-Tyr, aminovaleric acid,    pyroglutaminic acid, alpha-aminoisobutyric acid, gamma-aminobutyric    acid, alpha-aminobutyric acid, alpha,gamma-aminobutyric acid,    pyridylalanine, α-naphthylalanine, β-naphthylalanine,    Ac-β-naphthylalanine, N^(ε)-picoloyl-lysine, 4-halo-Phenyl,    4-pyrolidylalanine, isonipecotic carboxylic acid, and analogs,    derivatives, and isomers thereof.

-   32. The method of any of paragraphs 1-31, wherein the ghrelin    receptor antagonist is selected from the group consisting of    Tyr-DTrp-DLys-Trp-DPhe-NH₂ (SEQ ID NO: 11),    Tyr-DTrp-Lys-Trp-DPhe-NH₂ (SEQ ID NO: 12),    His-DTrp-DLys-Trp-DPhe-NH₂ (SEQ ID NO: 13),    His-DTrp-DLys-Phe-DTrp-NH₂ (SEQ ID NO: 14),    His-DTrp-DArg-Trp-DPhe-NH₂ (SEQ ID NO: 15),    His-DTrp-DLys-Trp-DPhe-Lys-NH₂ (SEQ ID NO: 16),    DesaminoTyr-DTrp-Ala-Trp-DPhe-NH₂ (SEQ ID NO: 17),    DesaminoTyr-DTrp-DLys-Trp-DPhe-NH₂ (SEQ ID NO: 18),    DeaminoTyr-DTrp-Ser-Trp-DPhe-Lys-NH₂ (SEQ ID NO: 19),    DesaminoTyr-DTrp-Ser-Trp-DPhe-NH₂ (SEQ ID NO: 20),    His-DTrp-DTrp-Phe-Met-NH₂ (SEQ ID NO: 21), Tyr-DTrp-DTrp-Phe-Phe-NH₂    (SEQ ID NO: 22), Glyψ[CH₂NH]-DβNal-Ala-Trp-DPhe-Lys-NH₂ (SEQ ID NO:    23), Glyψ[CH₂NH]-DβNal-DLys-Trp-DPhe-Lys-NH₂ (SEQ ID NO: 24),    DAla-DβNal-DLys-DTrp-Phe-Lys-NH₂ (SEQ ID NO: 25),    His-DβNal-DLys-Trp-DPhe-Lys-NH₂ (SEQ ID NO: 26),    Ala-His-DTrp-DLys-Trp-DPhe-Lys-NH₂ (SEQ ID NO: 27),    Alaψ[CH₂NH]-DβNal-Ala-Trp-DPhe-Lys-NH₂ (SEQ ID NO: 28),    DβNal-Ala-Trp-DPhe-Ala-NH₂ (SEQ ID NO: 29),    DAla-DcyclohexylAla-Ala-Phe-DPhe-Nle-NH₂ (SEQ ID NO: 30),    DcyclohexylAla-Ala-Phe-DTrp-Lys-NH₂ (SEQ ID NO: 31),    DAla-DβNal-Ala-Thr-DThr-Lys-NH₂ (SEQ ID NO: 32),    DcyclohexylAla-Ala-Trp-DPhe-NH₂ (SEQ ID NO: 33),    DAla-DβNal-Ala-Ala-DAla-Lys-NH₂ (SEQ ID NO: 34),    DβNal-Ala-Trp-DPhe-Leu-NH₂ (SEQ ID NO: 35),    His-DTrp-Phe-Trp-DPhe-Lys-NH₂ (SEQ ID NO: 36),    DAla-DβNal-DAla-DTrp-Phe-Lys-NH₂ (SEQ ID NO: 37),    βAla-Trp-DAla-DTrp-Phe-NH₂ (SEQ ID NO: 38),    His-Trp-DAla-DTrp-Phe-LysNH₂ (SEQ ID NO: 39),    DLys-DβNal-Ala-Trp-DPhe-Lys-NH₂ (SEQ ID NO: 40),    DAla-DβNal-DLys-DTrp-Phe-Lys-NH₂ (SEQ ID NO: 41),    Tyr-DAla-Phe-Aib-NH₂ (SEQ ID NO: 42), Tyr-DAla-Sar-NMePhe-NH₂ (SEQ    ID NO: 43), αγAbu-DTrp-DTrp-Ser-NH₂ (SEQ ID NO: 44),    αγAbu-DTrp-DTrp-Lys-NH₂ (SEQ ID NO: 45), αγAbu-DTrp-DTrp-Orn-NH₂    (SEQ ID NO: 46), αAbu-DTrp-DTrp-Orn-NH₂ (SEQ ID NO: 47),    DThr-DαNal-DTrp-DPro-Arg-NH₂ (SEQ ID NO: 48),    DAla-Ala-DAla-DTrp-Phe-Lys-NH₂ (SEQ ID NO: 49),    Alaψ[CH₂NH]His-DTrp-Ala-Trp-DPhe-Lys-NH₂ (SEQ ID NO: 50),    Lys-DHis-DTrp-Phe-NH₂ (SEQ ID NO: 51), γAbu-DTrp-DTrp-Orn-NH₂ (SEQ    ID NO: 52), inip-Trp-Trp-Phe-NH₂ (SEQ ID NO: 53),    Ac-DTrp-Phe-DTrp-Leu-NH₂ (SEQ ID NO: 54), Ac-DTrp-Phe-DTrp-Lys-NH₂    (SEQ ID NO: 55), Ac-DTrp-DTrp-Lys-NH₂ (SEQ ID NO: 56),    DLys-Tyr-DTrp-DTrp-Phe-Lys-NH₂ (SEQ ID NO: 57), Ac-DβNal-Leu-Pro-NH₂    (SEQ ID NO: 58), βAla-Trp-DTrp-DTrp-Orn-NH₂ (SEQ ID NO: 59),    DVal-DαNal-DTrp-Phe-Arg-NH₂ (SEQ ID NO: 60),    DLeu-DαNal-DTrp-Phe-Arg-NH₂ (SEQ ID NO: 61),    CyclohexylAla-DαNal-DTrp-Phe-Arg-NH₂ (SEQ ID NO: 62),    DTrp-DαNal-DTrp-Phe-Arg-NH₂ (SEQ ID NO: 63),    DAla-DβNal-DPro-Phe-Arg-NH₂ (SEQ ID NO: 64),    Ac-DαNal-DTrp-Phe-Arg-NH₂ (SEQ ID NO: 65), DαNal-DTrp-Phe-Arg-NH₂    (SEQ ID NO: 66), His-DTrp-DTrp-Lys-NH₂ (SEQ ID NO: 67),    Ac-DβNal-DTrp-NH₂ (SEQ ID NO: 68), αAib-DTrp-DcyclohexylAla-NH₂ (SEQ    ID NO: 69), αAib-DTrp-DAla-cyclohexylAla-NH₂ (SEQ ID NO: 70),    DAla-DcyclohexylAla-Ala-Ala-Phe-DPhe-Nle-NH₂ (SEQ ID NO: 71),    DPhe-Ala-Phe-DPal-NH₂ (SEQ ID NO: 72), DPhe-Ala-Phe-DPhe-Lys-NH₂    (SEQ ID NO: 73), DLys-Tyr-DTrp-DTrp-Phe-NH₂ (SEQ ID NO: 74),    Ac-DLys-Tyr-DTrp-DTrp-Phe-NH₂ (SEQ ID NO: 75),    Arg-DTrp-Leu-Tyr-Trp-Pro(cyclic Arg-Pro) (SEQ ID NO: 76),    Ac-DβNal-PicLys-ILys-DPhe-NH₂ (SEQ ID NO: 77),    DPal-Phe-DTrp-Phe-Met-NH₂ (SEQ ID NO: 78), DPhe-Trp-DPhe-Phe-Met-NH₂    (SEQ ID NO: 79), DPal-Trp-DPhe-Phe-Met-NH₂ (SEQ ID NO: 80),    βAla-Pal-DTrp-DTrp-Orn-NH₂ (SEQ ID NO: 81),    αγAbu-Trp-DTrp-DTrp-Orn-NH₂ (SEQ ID NO: 82),    βAla-Trp-DTrp-DTrp-Lys-NH₂ (SEQ ID NO: 83),    γAbu-Trp-DTrp-DTrp-Orn-NH₂ (SEQ ID NO: 84),    Ava-Trp-DTrp-DTrp-Orn-NH₂ (SEQ ID NO: 85),    DLys-Tyr-DTrp-Ala-Trp-DPhe-NH₂ (SEQ ID NO: 86),    His-DTrp-DArg-Trp-DPhe-NH₂ (SEQ ID NO: 87),    <Glu-His-Trp-DSer-DArg-NH₂ (SEQ ID NO: 88),    DPhe-DPhe-DTrp-Met-DLys-NH₂ (SEQ ID NO: 89),    Gly-Met-Ala-Gly-Ser-(Dap-Oct)-Phe-Leu-Ser-Pro-Glu-His-NH₂ (SEQ ID    NO: 3), O-(2-methylallyl)benzophonone oxime,    (R)-2-amino-3-(1H-indol-3-yl)-1-(4-phenylpiperidin-1-yl)propan-1-one,    N—((R)-1-((R)-1-((S)-3-(1H-indol-3-yl)-1-oxo-1-(4-phenylpiperidin-1-yl)propan-2-ylamino)-6-amino-1-oxohexan-2-ylamino)-3-hydroxy-1-oxopropan-2-yl)benzamide,    (S)—N—((S)-3-(1H-indol-3-yl)-1-oxo-1-(4-phenylpiperidin-1-yl)propan-2-yl)-6-acetamido-2-((S)-2-amino-3-(benzyloxy)propanamido)hexanamide,    (S)—N—((R)-3-(1H-indol-3-yl)-1-oxo-1-(4-phenylpiperidin-1-yl)propan-2-yl)-2-((S)-2-acetamido-3-(benzyloxy)propanamido)-6-aminohexanamide,    (R)—N-3-(1H-indol-3-yl)-1-(4-(2-methoxyphenyl)piperidin-1-yl)-1-oxopropan-2-yl)-4-aminobutanamide,    (R)—N-3-(1H-indol-3-yl)-1-(4-(2-methoxyphenyl)piperidin-1-yl)-1-oxopropan-2-yl)-2-amino-2-methylpropanamide,    methyl 3-(p-tolylcarbamoyl)-2-naphthoate, ethyl    3-(4-(2-methoxyphenyl)piperidine-1-carbonyl)-2-naphthoate,    3-(2-methoxyphenylcarbamoyl)-2-naphthoate,    (S)-2,4-diamino-N—((R)-3-(naphthalen-2-ylmethoxy)-1-oxo-1-(4-phenylpiperidin-1-yl)propan-2-yl)butanamide,    naphthalene-2,3-diylbis((4-(2-methoxyphenyl)piperazin-1-yl)methanone),    (R)-2-amino-N-(3-(benzyloxy)-1-oxo-1-(4-phenylpiperazin-1-yl)propan-2-yl)-2-methylpropanamide,    or (R)-2-amino-3-(benzyloxy)-1-(4-phenylpiperazin-1-yl)propan-1-one,    and pharmaceutically acceptable salts, prodrugs, or active    metabolites thereof.

-   33. The method of any of paragraphs 1-32, wherein the subject is a    mammal.

-   34. The method of any of paragraphs 1-33, wherein the subject is    non-human.

-   35. The method of any of paragraphs 1-34, wherein the subject is a    mouse or rat.

-   36. The method of any of paragraphs 1-32, wherein the subject is    human.

-   37. The method of any of paragraphs 1-35, wherein the GOAT inhibitor    and/or the ghrelin receptor antagonist is formulated in a    pharmaceutically acceptable formulation, wherein the formulation    comprises a pharmaceutically acceptable carrier.

-   38. The method paragraph 35, wherein the formulation is a sustained    release formulation.

-   39. The method of any of paragraphs 1-36, wherein said    administration is oral, parenteral, intravenous, intramuscular,    subcutaneous, transdermal, airway, pulmonary, nasal, rectal, buccal,    sublingual, or depot.

-   40. The method of any of paragraphs 1-38, wherein the GOAT inhibitor    and/or the ghrelin receptor antagonist is delivered continuously to    the subject.

-   41. The method of any of paragraphs 1-38, wherein the GOAT inhibitor    and/or the ghrelin receptor antagonist is delivered intermittently    to the subject

-   42. A peptide comprising the amino acid sequence of formula (VII):    A¹¹-A¹²-A¹³-Gly-Ser-A¹⁴-Phe-Leu-A¹⁵-A¹⁶-A¹⁷-A¹⁸ (SEQ ID NO: 93),    wherein each of A¹¹, A¹², and A¹³, is absent, an amino acid, or an    amino protecting group, and can be the same or different; each of    A¹⁵, A¹⁶, A¹⁷, and A¹⁸ is absent or an amino acid, and can be the    same or different; A¹⁴ is a serine conjugated with a —C(O)C₁-C₂₀    alky group on the side chain OH of said serine or a diaminopropionic    acid conjugated with a —C(O)C₁-C₂₀ alky group on one of the amino    group diaminopropionic acid, provided at least one of A¹¹, A¹², or    A¹³ is present.

-   43. The peptide of paragraph 41, wherein the peptide is a GOAT    inhibitor and/or a ghrelin receptor antagonist.

-   44. The peptide of any of paragraphs 41-42, wherein the amino acid    is selected from the group consisting of alanine, arginine,    asparagine, aspartic acid, cysteine, glutamic acid, glutamine,    glycine, histidine, isoleucine, leucine, lysine, methionine,    phenylalanine, proline, serine, threonine, tryptophan, tyrosine,    valine, homocysteine, phosphoserine, phosphothreonine,    phosphotyrosine, hydroxyproline, gamma-carboxyglutamate; hippuric    acid, octahydroindole-2-carboxylic acid, statine,    1,2,3,4,-tetrahydroisoquinoline-3-carboxylic acid, penicillamine    (3-mercapto-D-valine), ornithine, citruline, alpha-methyl-alanine,    para-benzoylphenylalanine, para-aminophenylalanine,    p-fluorophenylalanine, phenylglycine, propargylglycine, sarcosine,    and tert-butylglycine), diaminobutyric acid,    7-hydroxy-tetrahydroisoquinoline carboxylic acid, naphthylalanine,    biphenylalanine, cyclohexylalanine, amino-isobutyric acid,    norvaline, norleucine, tert-leucine, tetrahydroisoquinoline    carboxylic acid, pipecolic acid, phenylglycine, homophenylalanine,    cyclohexylglycine, dehydroleucine, 2,2-diethylglycine,    1-amino-1-cyclopentanecarboxylic acid,    1-amino-1-cyclohexanecarboxylic acid, amino-benzoic acid,    amino-naphthoic acid, gamma-aminobutyric acid,    difluorophenylalanine, nipecotic acid, alpha-amino butyric acid,    thienyl-alanine, t-butylglycine, desamino-Tyr, aminovaleric acid,    pyroglutaminic acid, alpha-aminoisobutyric acid, gamma-aminobutyric    acid, alpha-aminobutyric acid, alpha,gamma-aminobutyric acid,    pyridylalanine, α-naphthylalanine, β-naphthylalanine,    Ac-β-naphthylalanine, N^(ε)-picoloyl-lysine, 4-halo-Phenyl,    4-pyrolidylalanine, isonipecotic carboxylic acid, and isomers,    analogs and derivatives thereof.

-   45. The peptide of any of paragraphs 41-43, wherein the alkyl is a    C₁-C₁₆alkyl.

-   46. The peptide of any of paragraphs 41-44, wherein A⁵ is an    octanoylated serine or an octyanolyatd diaminopropionic acid.

-   47. The peptide of any of paragraphs 41-45, wherein the peptide    comprises at D-amino acid.

-   48. The peptide of any of paragraphs 41-46, wherein the peptide    comprises at least one beta-amino acid.

-   49. The peptide of any of paragraphs 41-47, wherein the peptide    comprises at least one peptide bond replaced by a linkage selected    from the group consisting of reduced psi peptide bond, urea,    thiourea, carbamate, sulfonyl urea, trifluoroethylamine,    ortho-(aminoalkyl)-phenylacetic acid, para-(aminoalkyl)-phenylacetic    acid, meta-(aminoalkyl)-phenylacetic acid, thioamide, tetrazole,    boronic ester, and olefinic group.

-   50. The peptide of any of paragraphs 41-48, wherein the peptide is    Gly-Met-Ala-Gly-Ser-(Dap-Oct)-Phe-Leu-Ser-Pro-Glu-His-NH₂ (SEQ ID    NO: 3).

Unless otherwise defined herein, scientific and technical terms used inconnection with the present application shall have the meanings that arecommonly understood by those of ordinary skill in the art.

It should be understood that this invention is not limited to theparticular methodology, protocols, and reagents, etc., described hereinand as such may vary. The terminology used herein is for the purpose ofdescribing particular embodiments only, and is not intended to limit thescope of the present invention, which is defined solely by the claims.

It should be understood that this invention is not limited to theparticular methodology, protocols, and reagents, etc., described hereinand as such may vary. The terminology used herein is for the purpose ofdescribing particular embodiments only, and is not intended to limit thescope of the present invention, which is defined solely by the claims.

Other than in the operating examples, or where otherwise indicated, allnumbers expressing quantities of ingredients or reaction conditions usedherein should be understood as modified in all instances by the term“about.” The term “about” when used in connection with percentages maymean±1%.

The invention is further illustrated by the following examples whichshould not be construed as limiting. The contents of all referencescited throughout this application, including the U.S. provisionalapplication Ser. No. 60/795,960 filed Apr. 28, 2006 as well as thefigures and tables are incorporated herein by reference.

Examples Animals and Experimental Procedures

Adult male Sprague Dawley rats (225-260 g) were purchased from CharlesRiver Canada (St. Constant, Canada) and individually housed on a 12-hlight, 12-h dark cycle (lights on, 0600-1800 h) in a temperature (22±1°C.)- and humidity-controlled room. Purina rat chow (Ralston Purina Co.,St. Louis, Mo.) and tap water were available ad libitum.

Chronic intracerebroventricular (i.c.v.) and intracardiac venouscannulas were implanted under sodium pentobarbital (50 mg/kg, i.p.)anesthesia using previously described techniques in (2) and (11). Theplacement of the icy cannulae was verified by both a positive drinkingresponse to i.c.v. carbachol (100 ng/10 μl) injection on the day aftersurgery and methylene blue dye at the time of death. After surgery, therats were placed directly in isolation test chambers with food and H₂Ofreely available until body weight returned to preoperative levels(usually within 5-7 d). During this time the rats were handled daily tominimize any stress associated with handling on the day of theexperiment. On the test day, food was removed 1.5 h before the start ofsampling and was returned at the end.

The efficacy of GHS-A to antagonize ghrelin's stimulatory action on GHat both central and peripheral sites was examined. For the centralexperiments, free-moving rats were i.c.v. injected with either normalsaline or GHS-A (5 μg) at 1045 h and, 15 min later (at 1100 h), weresubsequently administered ghrelin i.c.v. (500 ng). Both the humanghrelin peptide (provided by Dr. K. Chang, Phoenix Pharmaceuticals,Inc., Belmont, Calif.) and the GHS-A were diluted in normal saline justbefore use. Blood samples (0.35 ml) were withdrawn every 15 min over a6-h sampling period (1000-1600 h) from all animals. To document therapidity of the GH response to ghrelin, an additional blood sample wasobtained 5 min after injection of ghrelin. All blood samples wereimmediately centrifuged, and plasma was separated and stored at −20° C.for subsequent assay of GH. To avoid hemodynamic disturbance, the redblood cells were resuspended in normal saline and returned to the animalafter removal of the next blood sample.

For the peripheral experiments, free-moving rats were iv injected witheither normal saline or GHS-A (250 μg) at 1045 h and subsequently withghrelin i.v. (5 μg) at 1100 h. Blood samples were withdrawn from1000-1600 h, as described above.

For the study designed to assess the involvement of endogenous ghrelinin the genesis of pulsatile GH secretion, free-moving animals werei.c.v.-injected with either normal saline or GHS-A (5 μg) at twodifferent times in the 6-h sampling period: 1045 h and 1345 h. Thesetime points were chosen because they correspond closely to the time ofonset of the spontaneous GH secretory episodes, as previously documentedin our laboratory (2, 4). Blood samples were withdrawn from 1000-1600 h,as described above; however, no blood sample was withdrawn 5 min afterthe injections.

The effects of the GHS-A on both spontaneous and ghrelin-induced foodintake and body weight gain were investigated. For the spontaneousexperiments, the rats were fasted overnight (1600-1100 h next morning)and were i.c.v.-injected with either normal saline or GHS-A (5 μg) at1100 h. Food intake was monitored on an hourly basis for 5 h after theinitial injection (until 1600 h) and subsequently overnight (1600-0900 hnext morning). A measured amount of rat chow pellets was placed in thecage every hour. Spillage was collected by placement of a diaper underthe rat cages, and total food consumed for each period was calculated bysubtracting uneaten food plus spillage from total given. Body weightswere recorded daily at 0900 h. The latency to the onset of the firstmeal after the injection and the duration of that meal were alsomonitored.

To examine the effect of GHS-A on ghrelin-stimulated food intake,animals were icy injected with either normal saline or GHS-A (5 μg) at1045 h and subsequently with ghrelin (500 ng) at 1100 h. Food intake wasmonitored on an hourly basis as described above. In this experiment,food was removed 1.5 h before the start of the test.

All animal-based procedures were approved by the McGill UniversityAnimal Care Committee.

Receptor Binding and Calcium Mobilization Studies

The human ghrelin receptor type 1a (GHS-R1a) was expressed in HEK-293cells, whose cell membranes were subsequently harvested and used in thebinding assay. The receptor concentration (Bmax) used in the assay was2.3 pmol/mg of protein, resulting in a Kd for ghrelin binding of 0.016nM. The ability of the antagonist to displace 0.009 nM radiolabelledghrelin was then tested, at a concentration range of 0.1 nM to 10 μM.The binding affinities (Ki) for ghrelin, GHRP-6 and hexarelin in thissystem were 0.016 nM, 0.58 nM and 0.59 nM respectively.

The ability of the antagonist to mobilize calcium or to inhibitghrelin-stimulated calcium mobilization was examined using Euroscreen'sAequoScreen platform. This method is based on the co-expression inrecombinant cell lines of the GHS-R1a and aequorin, a photoproteincapable of detecting calcium concentrations in the lower micromolarrange. The agonistic properties of GHS-A were tested at a concentrationrange of 1 nM to 3 μM, and its capacity to inhibit the calcium mobilizedby 22.15 nM ghrelin was tested at a concentration range of 1 nM to 1.5μM. In this system, ghrelin was found to have an EC50 of 9.33 nM, EC80of 22.15 nM, and induced maximal activation at a concentration of 100nM.

The peptides were synthesized by the solid phase method and purified byHPLC. GH was determined by Radioimmunoassay (RIA).

The In Vitro Cell Culture Method

In vitro GH Release-Female rats of the CD-1 strain were housed in aconstant temperature room at 24° C. with 14 h light and 10 h darkness.The rats were fed Purina Rat chow and water at libitum. All studies werestarted between 0800-1000 hours. Pituitaries of mature female SpragueDawley rats were rapidly removed after decapitation, neurointermediatelobe discarded and then placed in a pH 7.4 buffer. The pituitaries werecut into ˜3-mm pieces and then transferred to a flask containing HEPSbuffer with trypsin and incubated at 37° C. Cells were trituratedseveral times during this period. After dispersion, the cells werecollected by centrifugation, wash with DMEM and placed into culturewell. Cell cultures were maintained for 4 days at 37° with 8% CO₂ addedto the incubator atmosphere. After 4 days in culture, cells were washedwith lactated Ringer's solution adjusted to pH 7.2-7.4 and then vehicle,peptide alone or peptide plus stimulator was added to media. Incubationtime was 60-120 minutes after which media was removed from each well forGH determination. The GH RIA reagents were distributed by the NIH.Control data was collected from cell cultures treated with only thecorresponding vehicle in the absence of peptide or stimulator. Controlstimulated data was collected from cell cultures treated with stimulatoralone in the absence of any peptide.

The In Vitro Pituitary Incubation Method

Hormonal activities were obtained from in vitro studies usingpituitaries of 20 day old CD-1 strain Sprague Dawley female rats. Twopituitaries were incubated for a total of 4-6 hours. Medium was removedeach hour for RIA of GH level and fresh medium was added again. Aftertwo one hour preincubation periods (P1-P2), the vehicle/peptides wereadded to two one hour incubations (I3-I4). Peptide activity wascalculated as the change in the hormonal level (delta) between I3+I4 andP2. For antagonist activity, the incubation was continued for 2additional hours (I5-I6) where both the peptide and a stimulator of GHsecretion was added and the antagonist activity was calculated as thechange in the hormonal level (delta) between I5+I6 and P2. The peptideswere assayed in triplicate and the hormone was assayed in duplicate.Each value recorded represents the mean of 6. The GH RIA reagents weredistributed by the NIH. Control data was collected from isolatedpituitary glands treated with only the corresponding vehicle in theabsence of peptide or stimulator. Control stimulated data was collectedfrom isolated pituitary glands treated with stimulator alone in theabsence of any peptide.

The In Vivo Assay

For the in vivo assay of GH Release in rat, immature female SpragueDawley rats were obtained from Charles River from Wilmington, Mass.After arrival they were housed at 25° C. with a 14:10 h light:darkcycle. Water and Purina rat chow were available at libitum. Pups wereweaned at 21 days of age.

Immature twenty six day old female Sprague Dawley rats, 3-6 rats pertreatment dose, were pretreated with pentobarbital 20 minutes before ivinjection of vehicle/peptide or peptide plus stimulator. Injection wasmade as a 0.1 ml solution. For the non-pentobarbital treated rat assay,peptides were administered iv into the tail vein of conscious rats. Allanimals were sacrificed by guillotine after iv peptide or vehicle. Trunkblood was collected at +10-15 minutes after decapitation, allowed toclot, centrifuged and serum stored until assayed for GH levels by RIA.The GH RIA reagents were distributed by the NIH. Control data wascollected from rats treated with only the corresponding vehicle in theabsence of peptide or stimulator. Control stimulated data was collectedfrom rats treated with stimulator alone in the absence of any peptide.

Results

The data obtained show that ghrelin receptor antagonists of the presentinvention such as HisDβNalDLysTrpDPheLysNH₂ (SEQ ID NO: 26), can be usedas a tool to disrupt the activity of ghrelin at the level of the CNS.This peptide is a GHRP derivative antagonist. ICV administration of 5 μgof this antagonist prior to i.c.v. injection of ghrelin (500 ng) in freemoving, adult rats virtually obliterated the GH response to ghrelin,FIG. 1. A similar block of ghrelin (5 μg iv) induced GH release wasobserved when rats were pretreated peripherally with the GHS-Rantagonist (250 μg i.v.), FIG. 2. In contrast, this GHS-R antagonist didnot significantly reduce the GH response to GHRH (5 μg i.v.). Withrespect to feeding, i.c.v. administered GHS antagonist (5 μg)significantly inhibited ghrelin's (500 ng i.c.v.) stimulatory effects onfood intake in the first hr after injections, FIGS. 7A and 8A. Theseresults show the modulatory role for endogenous ghrelin in maintainingthe high amplitude of spontaneous GH pulses under physiologicalconditions, likely acting through the GHS-R 1a on GHRH containingneurons in the arcuate nucleus (16, 17). Without wishing to be bound bytheory, while ghrelin may be necessary for the full response of GHRH(the major driving regulator) on pulsatile GH release, it is not anactive player in generating the ultradian rhythm of GH secretion. Thelack of a dissociated effect on GH and food intake by the GHS antagonistshows that the GHS-R 1a mediates the effects of ghrelin on feeding (viaNPY-containing neurons) as well as on GH.

The administration of the peptideGly-Met-Ala-Gly-Ser-(Dap-Oct)-Phe-Leu-Ser-Pro-Glu-His-NH₂ (SEQ ID NO: 3)reduced both cumulative food intake and inhibited plasma growth hormonelevels, FIGS. 10 and 11. Without wishing to be bound by theory, thispeptide acts on the hypothalamic ghrelin-R (GHS-R 1a) and inhibits theaction of endogenous ghrelin on its hypothalamic receptor. In thispeptide, the N-terminal, which is necessary for ghrelin's activity, ofthe ghrelin pentapeptide core is blocked by the tripeptide Gly-Met-Ala.

A similar peptide,Gly-Met-Ala-Gly-Ser-(Dap-Palmityl)-Phe-Leu-Ser-Pro-Glu-His-NH₂ (SEQ IDNO: 4), which has a Dap-palmityl residue instead of the Dap-Octanoyl didnot inhibit GH secretion or decreased food intake on administration,FIGS. 10 and 11. Without wishing to be bound by theory, the presence ofthe larger palmityl group may lower or inhibit the interaction of thispeptide with the ghrelin receptor.

The sequences disclosed in Tables 1-12 are as follows:Table 1 discloses SEQ ID NOS 96 and 11-22, respectively, in order ofappearance.Table 2 discloses SEQ ID NOS 97 and 23-28, respectively, in order ofappearance.Table 3 discloses SEQ ID NOS 29-39, respectively, in order ofappearance.Table 4 discloses SEQ ID NOS 97, 40, 40, 41, 41, 42, 42, 43 and 43,respectively, in order of appearance.Table 5 discloses SEQ ID NOS 44-53, respectively, in order ofappearance.Table 6 discloses SEQ ID NOS 97, 54, 54, 55, 55, 56, 56, 57, 57, 58, 58,59 and 59, respectively, in order of appearance.Table 7 discloses SEQ ID NOS 97, 60, 60, 61, 61-65, 65, 66, 66, 53 and53, respectively, in order of appearance.Table 8 discloses SEQ ID NOS 67-80, respectively, in order ofappearance.Table 9 discloses SEQ ID NOS 97 and 81-85, respectively, in order ofappearance.Table 10 discloses SEQ ID NO: 97.Table 11 discloses SEQ ID NO: 97.Table 12 discloses SEQ ID NOS 96, 98, 13, 27 and 86-89, respectively, inorder of appearance.

TABLE 1 In Vitro Ghrelin/GHRP/GHS Receptor Antagonist Activity onStimulated GH (ng/ml) release from isolated pituitary glands by thepituitary incubation method. Stimulator is His-DTrp-Ala-Trp-DPhe-Lys-NH₂at 10 ng/ml Stimulated Peptide Antagonist Dosage μg/ml PeptideAntagonist Control Control 0.1 0.3 1 1-Tyr-DTrp-DLys-Trp-DPhe-NH₂ 47 ±22 1528 ± 214 2-Tyr-DTrp-Lys-Trp-DPhe-NH₂ −461 ± 163  1053 ± 1823-His-DTrp-DLys-Trp-DPhe-NH₂ 57 ± 77 2120 ± 311 1765 ± 1604-His-DTrp-DLys-Phe-DTrp-NH₂ N/A 5-His-DTrp-DArg-Trp-DPhe-NH₂ −461 ±163  1953 ± 182 6-His-DTrp-DLys-Trp-DPhe-Lys-NH₂ −129 ± 52  1267 ± 64  952 ± 200 324 ± 181 7-DesaminoTyr-DTrp-Ala-Trp-DPhe-NH₂ 58 ± 77 2120 ±311 8-DesaminoTyr-DTrp-DLys-Trp-DPhe-NH₂ 223 ± 203  5189 ± 1513 4297 ±1061 2404 ± 802 688 ± 327 9-DeaminoTyr-DTrp-Ser-Trp-DPhe-Lys-NH₂  8 ±305  4436 ± 1006 10-DesaminoTyr-DTrp-Ser-Trp-DPhe-NH₂  8 ± 305  4436 ±1006 11-His-DTrp-DTrp-Phe-Met-NH₂ −129 ± 52  1267 ± 164 1542 ± 523 323 ±69  12-Tyr-DTrp-DTrp-Phe-Phe-NH₂ 47 ± 22 1528 ± 214 1274 ± 329 PeptideAntagonist Dosage μg/ml Peptide Antagonist 3 10 30 1001-Tyr-DTrp-DLys-Trp-DPhe-NH₂ −480 ± 95  −363 ± 662-Tyr-DTrp-Lys-Trp-DPhe-NH₂ −555 ± 121 3-His-DTrp-DLys-Trp-DPhe-NH₂ 949± 178    91 ± 103 4-His-DTrp-DLys-Phe-DTrp-NH₂5-His-DTrp-DArg-Trp-DPhe-NH₂ 341 ± 222 −125 ± 101 −122 ± 446-His-DTrp-DLys-Trp-DPhe-Lys-NH₂ 134 ± 91   −83 ± 132 −175 ± 597-DesaminoTyr-DTrp-Ala-Trp-DPhe-NH₂ 1302 ± 269  −959 ± 758-DesaminoTyr-DTrp-DLys-Trp-DPhe-NH₂ −466 ± 432  −1068 ± 318   −576 ±110 9-DeaminoTyr-DTrp-Ser-Trp-DPhe-Lys-NH₂ 3325 ± 391  3810 ± 62110-DesaminoTyr-DTrp-Ser-Trp-DPhe-NH₂ 3119 ± 488  3258 ± 68211-His-DTrp-DTrp-Phe-Met-NH₂ 445 ± 188 287 ± 68 −319 ± 9512-Tyr-DTrp-DTrp-Phe-Phe-NH₂ 1034 ± 182  −167 ± 157

TABLE 2 In Vivo Ghrelin/GHRP/GHS Receptor Antagonist Activity onStimulated GH (ng/ml) release from rat. Sitmulator isDAla-DβNal-Ala-Trp-DPhe-Lys-NH2 at 0.3 μg or 1 μg Stimulated StimulatedControl Control Peptide Antagonist Dosage μg/i.v. Peptide AntagonistControl 0.3 μg 1 μg 1 3 10 30 100 1-Glyψ[CH₂NH]-DβNal-Ala- 138 ± 11 1412± 400 1112 ± 200 578 ± 82 1198 ± 114 Trp-DPhe-Lys-NH₂ 138 ± 11 3214 ±276 2307 ± 176 890 ± 236 164 ± 14 3105 ± 429 1842 ± 454 1135 ± 1402-Glyψ[CH₂NH]-DβNal-DLys- 143 ± 19 2406 ± 288 2305 ± 320 1990 ± 196 1550± 284 946 ± 133 462 ± 122 Trp-DPhe-Lys-NH₂ 3-DAla-DβNal-DLys-DTrp-Phe-327 ± 39 4950 ± 98  2884 ± 828 1198 ± 114 Lys-NH₂4-His-DβNal-DLys-Trp-DPhe-  91 ± 46 2253 ± 252 733 ± 85 Lys-NH₂  91 ± 462825 ± 134 818 ± 269 5-Ala-His-DTrp-DLys-Trp-DPhe-  91 ± 46 2253 ± 2521487 ± 397 Lys-NH₂ 6-Alaψ[CH₂NH]-DβNal-Ala-Trp- 164 ± 14 3104 ± 429 2771± 157 2341 ± 416 1948 ± 450 1639 ± 221 DPhe-Lys-NH₂

TABLE 3 In Vivo Ghrelin/GHRP/GHS Receptor Antagonist Activity onUnstimulated Basal GH (ng/ml) Release from rat. Stimulated StimulatedControl Control Peptide Dosage μg/i.v. Partial Agonist/AntagonistPeptide Control 0.3 μg 1 μg 1 3 10 30 100 1-DβNal-Ala-Trp-DPhe-Ala-NH₂253 ± 34 1991 ± 214 623 ± 60 694 ± 70 654 ± 58 713 ± 712-DAla-DcyclohexylAla-Ala- 204 ± 46 1850 ± 324  435 ± 152 195 ± 34 250 ±41 393 ± 51 697 ± 75 Phe-DPhe-Nle-NH₂ 3-DcyclohexylAla-Ala- 204 ± 461765 ± 330 199 ± 63 266 ± 68 199 ± 23 346 ± 82 350 ± 61 Phe-DTrp-Lys-NH₂4-DAla-DβNal-Ala-Thr- 244 ± 56 1538 ± 215 255 ± 38 288 ± 31 386 ± 57DThr-Lys-NH₂ 5-DcyclohexylAla-Ala- 176 ± 44 2282 ± 258 181 ± 28 237 ± 18354 ± 81 771 ± 76 Trp-DPhe-NH₂ 6-DAla-DβNal-Ala- 135 ± 19 1485 ± 200 235± 43 178 ± 33 172 ± 15 185 ± 39 Ala-DAla-Lys-NH₂7-DβNal-Ala-Trp-DPhe-Leu-NH₂ 145 ± 48 1470 ± 338 253 ± 79 277 ± 43 347 ±66  645 ± 117 8-His-DTrp-Phe-Trp-DPhe-Lys-NH₂ 240 ± 55 2766 ± 726  67 ±14 141 ± 53 197 ± 70 509 ± 48 9-DAla-DβNal-DAla- 100 ± 22 4785 ± 798 184± 55  467 ± 201  244 ± 107 DTrp-Phe-Lys-NH₂10-βAla-Trp-DAla-DTrp-Phe-NH₂ 195 ± 33 4130 ± 349 341 ± 46  636 ± 17111-His-Trp-DAla-DTrp-Phe-LysNH₂ 150 ± 26 1847 ± 362 204 ± 44 127 ± 44 83± 5

TABLE 4 In Vivo Ghrelin/GHRP/GHS Receptor Antagonist Activity onUnstimulated and Stimulated (S) GH (ng/ml) release from rats. TheStimulator is DAla-DβNal-Ala-Trp-DPhe-Lys-NH2 at 0.3 μg, 1 μg, or 10 μg.Stimulated Stimulated Control Control Peptide Antagonist Dosage μg/i.v.Peptide Antagonist (P) Control 0.3 1 1 3 10 30 100 Pentobarbital Rats1-DLys-DβNal-Ala-Trp- −S 197 ± 81  616 ± 169 847 ± 17  629 ± 146 228 ±45 DPhe-Lys-NH₂ 1-DLys-DβNal-Ala-Trp- P + S 5052 ± 511 5232 ± 346 3404 ±396  704 ± 169 DPhe-Lys-NH₂ 2-DAla-DβNal-DLys-DTrp- −S 327 ± 39 323 ± 50812 ± 6  Phe-Lys-NH₂ 2-DAla-DβNal-DLys-DTrp- P + S 4950 ± 98  2864 ± 8281198 ± 114 Phe-Lys-NH₂ Non-Pentobarbital Rats 10 μg1-Tyr-DAla-Phe-Alb-NH₂ −S 12 ± 1 18 ± 1 1-Tyr-DAla-Phe-Alb-NH₂ P + S 72± 9 23 ± 5 2-Tyr-DAla-Sar-NMePhe-NH₂ −S 12 ± 1 18 ± 42-Tyr-DAla-Sar-NMePhe-NH₂ P + S 72 ± 9 24 ± 6

TABLE 5 In Vivo Ghrelin/GHRP/GHS Receptor Antagonist Activity onUnstimulated Basal GH (ng/ml) release from rats. Stimulated StimulatedControl Control Peptide Antagonist Dosage μg/i.v. Peptide AntagonistControl 0.3 μg 1 μg 1 3 10 30 100 1-αγAbu-DTrp-DTrp-Ser-NH₂ 106 ± 9 2742 ± 206  80 ± 35  62 ± 15 67 ± 8 2-αγAbu-DTrp-DTrp-Lys-NH₂ 136 ± 311968 ± 294 57 ± 7  84 ± 18  62 ± 15 3-αγAbu-DTrp-DTrp-Orn-NH₂ 167 ± 132819 ± 530 118 ± 16 126 ± 27  79 ± 31 4-αAbu-DTrp-DTrp-Orn-NH₂ 167 ± 132819 ± 530  85 ± 25  88 ± 18 50 ± 6 5-DThr-DαNal-DTrp-DPro-Arg-NH₂ 164 ±23 2691 ± 281 60 ± 5 130 ± 24 134 ± 31 6-DAla-Ala-DAla-DTrp-Phe-Lys-NH₂180 ± 20 4785 ± 798 228 ± 76 172 ± 14 153 ± 457-Alaψ[CH₂NH]His-DTrp-Ala-Trp-DPhe-Lys-NH₂ 211 ± 30 2335 ± 323 127 ± 32147 ± 37 8-Lys-DHis-DTrp-Phe-NH₂ 211 ± 30 2335 ± 323 121 ± 249-γAbu-DTrp-DTrp-Orn-NH₂ 167 ± 13 2819 ± 530  82 ± 28 90 ± 5 113 ± 3210-inip-Trp-Trp-Phe-NH₂ 155 ± 31 2503 ± 240 69 ± 3  81 ± 10 inip =isonipecotic carboxylic acid αγAbu = alpha gamma diaminobutyric acid

TABLE 6 In Vitro Ghrelin/GHRP/GHS Receptor Antagonist Activity onUnstimulated and Stimulated(S) GH (ng/ml) release from cell culture ofpituitary cells. The Sitmulator is DAla-DβNal-Ala-Trp-DPhe-Lys-NH₂ at at10 ng/ml. Stimulated Control Peptide Antagonist Dosage μg/m/l PeptideAntagonist (P) Control 10 ng/ml 0.1 0.3 1 3 10 301-Ac-DTrp-Phe-DTrp-Leu-NH₂ −S 1640 ± 100 400 ± 201-Ac-DTrp-Phe-DTrp-Leu-NH₂ P + S 2420 ± 0 2100 ± 0  1200 ± 20 600 ± 202-Ac-DTrp-Phe-DTrp-Lys-NH₂ −S 1640 ± 100 350 ± 802-Ac-DTrp-Phe-DTrp-Lys-NH₂ P + S 2420 ± 0 1750 ± 10 800 ± 0 470 ± 303-Ac-DTrp-DTrp-Lys-NH₂ −S 1640 ± 100  610 ± 30 420 ± 203-Ac-DTrp-DTrp-Lys-NH₂ P + S 2420 ± 0 1970 ± 70 1130 ± 30 900 ± 0 4-DLys-Tyr-DTrp-DTrp-Phe-Lys-NH₂ −S 1640 ± 100 1340 ± 60 1060 ± 0 4-DLys-Tyr-DTrp-DTrp-Phe-Lys-NH₂ P + S 2420 ± 0 2100 ± 40 1710 ± 10 1270± 10  5-Ac-DβNal-Leu-Pro-NH₂ −S 1233 ± 49  5-Ac-DβNal-Leu-Pro-NH₂ P + S 2811 ± 229 1998 ± 36 1206 ± 53 860 ± 33 6-βAla-Trp-DTrp-DTrp-Orn-NH₂ −S1722 ± 205 6-βAla-Trp-DTrp-DTrp-Orn-NH₂ P + S 2385 ± 8 3103 ± 471 1633 ±34 1166 ± 13 

TABLE 7 In Vitro Ghrelin/GHRP/GHS Receptor Antagonist Activity onUnstimulated and Stimulated (S) GH (ng/ml) release from cell culture ofpituitary cells. The Stimulator is DAla-DβNal-Ala-Trp-DPhe-Lys-NH₂ at 1ng/ml. Stimulated Control Peptide Dosage μg/ml PartialAgonist/Antagonist Peptide (P) Control 1 ng/ml 1 3 10 30 1001-DVal-DαNal-DTrp-Phe-Arg-NH₂ −S 480 ± 16  934 ± 34 850 ± 19 598 ± 7 1-DVal-DαNal-DTrp-Phe-Arg-NH₂ P + S 1399 ± 27  949 ± 52 672 ± 64 520 ±5  2-DLeu-DαNal-DTrp-Phe-Arg-NH₂ −S 480 ± 16 1156 ± 10 971 ± 5  520 ± 5 2-DLeu-DαNal-DTrp-Phe-Arg-NH₂ P + S 1399 ± 27 1136 ± 7  957 ± 44 777 ±71 3-CyclohexylAla-DαNal-DTrp-Phe-Arg-NH₂ P + S 734 ± 6  1841 ± 41 1362± 59 1021 ± 22  4-DTrp-DαNal-DTrp-Phe-Arg-NH₂ P + S 734 ± 6  1851 ± 411239 ± 17 878 ± 28 5-DAla-DβNal-DPro-Phe-Arg-NH₂ P + S 734 ± 6  1851 ±41 1779 ± 27 1328 ± 59  6-Ac-DαNal-DTrp-Phe-Arg-NH₂ −S 480 ± 16 1106 ±7  996 ± 16 704 ± 76 6-Ac-DαNal-DTrp-Phe-Arg-NH₂ P + S 1399 ± 27 1128 ±12 970 ± 25 704 ± 76 7-DαNal-DTrp-Phe-Arg-NH₂ −S 480 ± 16 1170 ± 43 987± 52 727 ± 44 7-DαNal-DTrp-Phe-Arg-NH₂ P + S 1399 ± 27 1288 ± 40 1079 ±17  824 ± 29 8-inip-Trp-Trp-Phe-NH₂ −S 625 ± 12  553 ± 111 247 ± 9  132± 7 8-inip-Trp-Trp-Phe-NH₂ P + S  749 ± 28 393 ± 6  278 ± 35 154 ± 4inip = isonipecotic carboxylic acid

TABLE 8 In Vitro Ghrelin/GHRP/GHS Receptor Antagonist Activity onUnstimulated Basal GH (ng/ml) release from cell cultures of pituitarycells. Stimulated control Peptide Antagonist Dosage μg/ml PeptideAntagonist (P) Control 1 ng/ml 1 3 10 30 100 1-His-DTrp-DTrp-Lys-NH₂1089 ± 47 1551 ± 2 1124 ± 37 749 ± 10 615 ± 41 2-Ac-DβNal-DTrp-NH₂ 1089± 47 1551 ± 2 1264 ± 2  980 ± 72 699 ± 7  3-αAib-DTrp-DcyclohexylAla-NH₂478 ± 8 1014 ± 8  980 ± 44  826 ± 32 602 ± 53 492 ± 114-αAib-DTrp-DAla-cyclohexylAla-NH₂ 478 ± 8 1014 ± 8 1086 ± 52 1103 ± 18994 ± 22  704 ± 115 5-DAla-DcyclohexylAla-Ala-Ala-  500 ± 116  1991 ±214  286 ± 75  177 ± 44 271 ± 38 376 ± 28 Phe-DPhe-Nle-NH₂6-DPhe-Ala-Phe-DPal-NH₂  176 ± 44  170 ± 19 181 ± 31 161 ± 20 146 ± 217-DPhe-Ala-Phe-DPhe-Lys-NH₂  368 ± 32 267 ± 27 276 ± 65 360 ± 848-DLys-Tyr-DTrp-DTrp-Phe-NH₂ 1403 ± 13 1451 ± 19 1175 ± 77 1129 ± 6  744± 44 9-Ac-DLys-Tyr-DTrp-DTrp-Phe-NH₂ 1403 ± 13 105 ± 8 950 ± 91 782 ± 56756 ± 1  10-Arg-DTrp-Leu-Tyr-Trp-Pro(cyclic Arg-Pro) 1403 ± 13 1480 ± 19802 ± 26 601 ± 16 509 ± 49 11-Ac-DβNal-PlcLys-ILys-DPhe-NH₂ 1333 ± 41 1013 ± 207 976 ± 13 928 ± 16 12-DPal-Phe-DTrp-Phe-Met-NH₂ 1333 ± 411081 ± 50 997 ± 30 425 ± 25 13-DPhe-Trp-DPhe-Phe-Met-NH₂ 1333 ± 41 1146± 34 1086 ± 32  871 ± 89 14-DPal-Trp-DPhe-Phe-Met-NH₂ 1333 ± 41 1105 ±18 891 ± 4  567 ± 24 ILys = Lys(IPr)

TABLE 9 In Vitro Ghrelin/GHRP/GHS Receptor Antagonist Activity onUnstimulated and Stimulated(S) GH (ng/ml) release from cell cultures ofpituitary cells. The Stimulator is DAla-DβNal-Ala-Trp-DPhe-Lys-NH₂ at 1ng/ml. Stimulated control Peptide Antagonist Dosage μg/ml PeptideAntagonist (P) Control 1 ng/ml 1 3 10 30 1001-βAla-Pal-DTrp-DTrp-Orn-NH₂ −S 894 ± 18 759 ± 11 861 ± 25 P + S 1232 ±34 855 ± 11 828 ± 11 2-αγAbu-Trp-DTrp-DTrp-Orn-NH₂ −S 894 ± 18 609 ± 3 503 ± 5  P + S 1232 ± 34 666 ± 2  578 ± 31 3-βAla-Trp-DTrp-DTrp-Lys-NH₂−S 894 ± 18 733 ± 25 616 ± 21 P + S 1232 ± 34 806 ± 45 596 ± 184-γAbu-Trp-DTrp-DTrp-Orn-NH₂ −S 894 ± 18 840 ± 30 634 ± 1  P + S 1232 ±34 835 ± 5  655 ± 40 5-Ava-Trp-DTrp-DTrp-Orn-NH₂ −S 894 ± 18 481 ± 3 406 ± 21 P + S 1232 ± 34 505 ± 19 420 ± 34 αγAbu = alpha gammadiaminobutyric acid Ava = aminovaleric acid

TABLE 10 In Vitro Ghrelin/GHRP/GHS Receptor Antagonist Activity onUnstimulated and Stimulated(S) GH (ng/ml) release from cell cultures ofpituitary cells. The Stimulator is DAla-DβNal-Ala-Trp-DPhe-Lys-NH₂ at 1ng/ml. Stimulated Control Partial Peptide/Non-peptide Dosage μg/mlPartial Peptide/Non-peptide(P) Control 1 ng/ml 1 3 10 301-DTrp-4-phenylpiperdinamide 385 ± 49 815 ± 26 390 ± 35 520 ± 61 577 ±24 P + S 1060 ± 24 1085 ± 1  917 ± 4  344 ± 6  486 ± 292-2,3-di[N-(2-methoxylphenyl) 361 ± 30 338 ± 3  204 ± 10 262 ± 4 piperazyl-naphthalene carboxylamide P + S 905 ± 6 654 ± 18 442 ± 4  537± 28 385 ± 17 136 ± 11 118 ± 8  P + S 1153 ± 36 648 ± 16 309 ± 463-Benzamide-DSerDLysTrp-p- 370 ± 24 393 ± 54 369 ± 30phenylpiperidinamide P + S 1216 ± 26 432 ± 25 353 ± 104-Ser(Bzl)Lys(Ac)DTrp-p- 385 ± 17 388 ± 41 273 ± 39 phenylpiperidinamideP + S 1153 ± 36 571 ± 32 399 ± 24 5-O-(2-methylallyl) benzophonone oxime969 ± 33 929 ± 28 616 ± 23 P + S 1461 ± 58 1281 ± 58  699 ± 53 6-DSer(BZL)-N′-phenyl-N-piperazinamide 626 ± 4  585 ± 10 368 ± 2  P + S1016 ± 18 719 ± 26 435 ± 0  7-αAibDSer(BZL)-N′-phenyl-N-piperazinamide626 ± 4  777 ± 34 499 ± 18 P + S 1016 ± 18 878 ± 30 510 ± 158-2-[acetylester]-3-(p-m-methoxyl phenyl) 421 ± 16 373 ± 2  176 ± 11piperidinamide]-naphthalene carboxamide P + S 859 ± 4 480 ± 9  223 ± 22

TABLE 11 In Vitro Ghrelin/GHRP/GHS Receptor Antagonist Activity onUnstimulated and Stimulated(S) GH (ng/ml) release from cell cultures ofpituitary cells. The Stimulator is DAla-DβNal-Ala-Trp-DPhe-Lys-NH₂ at 1ng/ml. Stimulated Control Partial Peptide/Non-peptide Dosage μg/mlPartial Peptide/Non-peptide(P) Control 1 ng/ml 1 3 10 301-2-[methylester]-3-[p-methylphenylamide 626 ± 4  754 ± 32 498 ± 26naphthalene carboxylamide P + S 1016 ± 18 1149 ± 33  886 ± 292-p-phenyl(piperidinamide-DTrpLysSer(BZL)- P + S 408 ± 40 905 ± 6 680 ±13 489 ± 41 245 ± 16 acetylamide 3-γAbuDTrp-p-[m-methoxyphenyl] 364 ± 31557 ± 19 378 ± 18 piperidinamide P + S  947 ± 11 526 ± 27 428 ± 224-αAibDTrp-p-(α-methoxylphenyl) 377 ± 24 365 ± 2  375 ± 30piperidinamide P + S  947 ± 33 441 ± 21 384 ± 165-2-[ethylester-3-m-methoxylphenylamide] 364 ± 31 698 ± 18 552 ± 20naphthalene carboxylamide P + S  947 ± 11 670 ± 32 458 ± 156-1,3-diaminobutyricamide-DβNal-4- 626 ± 4  794 ± 34 504 ± 20phenylpiperidinamide P + S 1016 ± 18 644 ± 33 529 ± 20

TABLE 12 In Vitro Ghrelin/GHRP/GHS Receptor Antagonist Activity onUnstimulated and Stimulated(S) GH (ng/ml) release from isolatedpituitary glands by the pituitary incubation method. The Stimulator S =His-DTrp-Ala- Trp-DPhe-Lys-NH₂ (10 ng/ml) and Stimulator *S =Tyr-DTrp-Ala-Trp-DPhe-NH₂ (0.3 μg/ml) Stimulated Peptide Dosage μg/mlPeptide(P) Control Control 1 3 10 30 100 1-His-DTrp-DLys-Trp-DPhe-NH₂P + S 854 ± 255 8769 ± 583 8121 ± 687 5929 ± 857 3017 ± 413   269 ± 1402-Ala-His-DTrp-DLys-Trp-DPhe-Lys-NH₂ 1674 ± 1171 3977 ± 1360 P + S 5218± 507 4850 ± 539  947 ± 551 −2384 ± 868 3-DLys-Tyr-DTrp-Ala-Trp-DPhe-NH₂* P + S 148 ± 137 2218 ± 194 1233 ± 268 688 ± 3233 916 ± 80 4-His-DTrp-DArg-Trp-DPhe-NH₂ P + S −14 ± 62  −109 ±124 −500 ± 104  776 ± 142  136 ± 108 290 ± 124 −454 ± 95 5-<Glu-His-Trp-DSer-DArg-NH₂ 246 ± 67  −4 ± 25  6 ± 346-DPhe-DPhe-DTrp-Met-DLys-NH₂* P + S 148 ± 137 2218 ± 194 1584 ± 1361398 ± 98  1388 ± 300

All patents and other publications identified throughout thespecification and examples and in the references section are expresslyincorporated herein by reference for the purpose of describing anddisclosing, for example, the methodologies described in suchpublications that might be used in connection with the presentinvention. These publications are provided solely for their disclosureprior to the filing date of the present application. Nothing in thisregard should be construed as an admission that the inventors are notentitled to antedate such disclosure by virtue of prior invention or forany other reason. All statements as to the date or representation as tothe contents of these documents is based on the information available tothe applicants and does not constitute any admission as to thecorrectness of the dates or contents of these documents.

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All references cited in the specification and the Examples areincorporated herein in their entirety by reference.

What is claimed is:
 1. A method for treatment, prevention or managementof obesity or obesity related disease or disorder in a subject in needthereof, said method comprising the step of administering an effectiveamount of a ghrelin O-acyltransferase (GOAT) inhibitor and an effectiveamount of a ghrelin receptor antagonist.